Substituted anilines as ccr(4) antagonists

ABSTRACT

Aniline compounds are provided which bind to CCR(4) and are useful for the treatment of diseases such as allergic diseases, autoimmune diseases, graft rejection and cancer.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.15/674,230 filed Aug. 10, 2017, which is a continuation of U.S. patentapplication Ser. No. 15/207,315 filed Jul. 11, 2016 (now U.S. Pat. No.9,758,512), which is a divisional of U.S. patent application Ser. No.14/452,341 filed Aug. 5, 2014 (now U.S. Pat. No. 9,567,323) which is acontinuation of U.S. patent application Ser. No. 13/691,571 filed Nov.30, 2012 (now U.S. Pat. No. 8,835,419) which application claims thebenefit of priority to U.S. Provisional Application Ser. No. 61/565,968,filed Dec. 1, 2011, each of which is incorporated herein by reference inits entirety.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

Not Applicable

REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

Not Applicable

BACKGROUND OF THE INVENTION

Chemokines are chemotactic cytokines that are released by a wide varietyof cells to attract macrophages, T cells, eosinophils, basophils andneutrophils to sites of inflammation (reviewed in Schall, Cytokine,3:165-183 (1991), Schall, et al., Curr. Opin. Immunol. 6:865-873 (1994)and Murphy, Rev. Immun., 12:593-633 (1994)). In addition to stimulatingchemotaxis, other changes can be selectively induced by chemokines inresponsive cells, including changes in cell shape, transient rises inthe concentration of intracellular free calcium ions ([Ca2+]), granuleexocytosis, integrin upregulation, formation of bioactive lipids (e.g.,leukotrienes) and respiratory burst, associated with leukocyteactivation. Thus, the chemokines are early triggers of the inflammatoryresponse, causing inflammatory mediator release, chemotaxis andextravasation to sites of infection or inflammation.

There are two main classes of chemokines, CXC (alpha) and CC (beta),depending on whether the first two cysteines are separated by a singleamino acid (C—X—C) or are adjacent (C—C). The alpha-chemokines, such asinterleukin-8 (IL-8), neutrophil-activating protein-2 (NAP-2) andmelanoma growth stimulatory activity protein (MGSA) are chemotacticprimarily for neutrophils, whereas beta-chemokines, such as RANTES,MIP-la, MIP-lb, monocyte chemotactic protein-1 (MCP-1), MCP-2, MCP-3 andeotaxin are chemotactic for macrophages, T-cells, eosinophils andbasophils (Deng, et al., Nature, 381:661-666 (1996)). The chemokinesbind specific cell-surface receptors belonging to the family ofG-protein-coupled seven-transmembrane-domain proteins (reviewed inHoruk, Trends Pharm. Sci., 15:159-165 (1994)) which are termed“chemokine receptors.”

On binding their cognate ligands, chemokine receptors transduce anintracellular signal though the associated trimeric G protein, resultingin a rapid increase in intracellular calcium concentration. There are atleast eleven human chemokine receptors that bind or respond tobeta-chemokines and at least seven human chemokine receptors that bindto the alpha chemokines. Additionally CX3CR1 (fractalkine receptor) canbind to the fractalkine chemokine, which is distinguished by a series ofthree amino acids between the first two cysteines. Chemokine receptors,have been implicated as being important mediators of inflammatory andimmunoregulatory disorders and diseases, including asthma and allergicdiseases, as well as autoimmune pathologies such as rheumatoid arthritisand atherosclerosis.

The CC Chemokine receptor 4, CCR(4), first identified by Power et al.(Power et al. (1995) J Biol. Chem. 270:19495-19500), is a Gprotein-coupled receptor that binds to chemokines including CCL22, alsoknown as Macrophage-Derived Chemokine (MDC; a CC chemokine reported tobe a chemoattractant for the Th2 subset of peripheral blood T cells,dendritic cells, and natural killer (NK) cells), and CCL17, also knownas TARC (thymus and activation-regulated chemokine), which is alsoproduced by monocytes and dendritic cells.

The full-length human CCR(4) protein (GenBank Accession No. X85740;SWISS-PROT Accession No. P51679) has been described, see, e.g, Imai etal. (1998) J. Biol. Chem. 273:1764-1768.

While the global distribution of CCR(4) is unknown, the receptor isexpressed primarily in peripheral blood T lymphocytes, and is found onapproximately 20% of adult peripheral blood effector/memory CD4+ Tcells. CCR(4) is involved in T lymphocyte homing to the skin and lungs(see, e.g., Campbell et al. (1999) Nature 400:776-780, Gonzalo et al.(1999) J. Immunol. 163:403-5 411, Lloyd et al. (2000) J. Exp. Med.191:265-273, Kawasaki et al. (2001) J. Immunol. 166:2055-2062) and isfound on almost all T cells that have a skin homing phenotype, the CTLA+T cells. Thus CCR(4) may be an important player in skin pathologies inwhich leukocytes participate. It also seems likely that CCR(4) isexpressed on some other cell types, probably monocytes/macrophages anddendritic cells, among others. In view of the clinical importance ofCCR(4), the identification of compounds that modulate CCR(4) functionrepresent an attractive avenue into the development of new therapeuticagents. Such compounds and methods for their use are provided herein.

BRIEF SUMMARY OF THE INVENTION

In the present disclosure, compounds are provided having formula (I):

wherein the groups/letters R¹, R², R³, R⁴, R⁵, R⁶, R⁷, R^(a), R^(b), B,Q, W, X, Y, Z having the meanings provided in the Detailed Description.

Compositions containing the compounds of formula (I) are provided aswell as methods for treating diseases and conditions modulated by CCR(4)activity. Still further, the compounds provided herein are useful inmethods of screening for additional CCR(4)-modulatory compounds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 provides three reaction schemes (eq. 1, eq. 2 and eq. 3) usefulin construction of portions of the compounds provided herein.

FIG. 2 provides seven reaction schemes (eq. 4, eq. 5, eq. 6, eq. 7, eq.8, eq. 9, and eq. 10) useful in construction of portions of thecompounds provided herein.

FIG. 3 provides four reaction schemes (eq. 11, eq. 12, eq. 13, and eq.14) useful in construction of portions of the compounds provided herein.

FIG. 4 provides four reaction schemes (eq. 15, eq. 16, eq. 17, and eq.18) useful in construction of portions of the compounds provided herein.

FIG. 5 provides a reaction scheme for the preparation of2-chloro-N-((1R)-1-(2,4-dichlorophenyl)ethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(see Example 2).

FIG. 6 provides a reaction scheme for the preparation of2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)-5-(1-((S)-3-piperidyl)-3,6-dihydro-2H-pyridin-4-yl)aniline(see Example 3).

FIG. 7 provides a reaction scheme for the preparation of2-((S)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)piperidin-1-yl)acetic acid (see Example 4).

FIG. 8 provides a reaction scheme for the preparation of3-((S)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)piperidin-1-yl)propanoic acid (see Example 5).

FIG. 9 provides a reaction scheme for the preparation of5-((S)-1,3′-bipiperidin-4-yl)-2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)aniline(see Example 6).

FIG. 10 provides a reaction scheme for the preparation of2-((S)-4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-1,3′-bipiperidin-1′-yl)aceticacid (see Example 7).

FIG. 11 provides a reaction scheme for the preparation of3-((S)-4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-1,3′-bipiperidin-1′-yl)propanoicacid (see Example 8).

FIG. 12 provides a reaction scheme for the preparation of(1R,3R)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-1-yl)cyclohexanecarboxamide(see Example 9).

FIG. 13 provides a reaction scheme for the preparation of(R)-(1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)(morpholino)methanone(see Example 10).

FIG. 14 provides a reaction scheme for the preparation of(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)((S)-3-methoxypyrrolidin-1-yl)methanone(see Example 11).

FIG. 15 provides a reaction scheme for the preparation of1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidine-4-carbonyl)pyrrolidine-3-carbonitrile(see Example 12).

FIG. 16 provides a reaction scheme for the preparation of(4-(3-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazin-1-yl)((R)-piperidin-2-yl)methanone(see Example 13).

FIG. 17 provides a reaction scheme for the preparation of(4-(3-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazin-1-yl)((R)-pyrrolidin-2-yl)methanone(see Example 14).

FIG. 18 provides a reaction scheme for the preparation of3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexanecarboxamide(see Example 15).

FIG. 19 provides a reaction scheme for the preparation of(3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexyl)methanol(see Example 16).

FIG. 20 provides a reaction scheme for the preparation of2-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-(4-((R)-piperidine-2-carbonyl)piperazin-1-yl)benzonitrile(see Example 17).

FIG. 21 provides a reaction scheme for the preparation of1-(1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)pyrrolidin-3-yl)urea(see Example 18).

FIGS. 22A-22S provide structure and activity for representativecompounds provided herein (see also Biological Example 1).

DETAILED DESCRIPTION OF THE INVENTION I. Abbreviation and Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group having one or more double bonds. Similarly,the term “alkynyl” refers to an unsaturated alkyl group having one ormore triple bonds. Examples of such unsaturated alkyl groups includevinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. “Cycloalkyl” is also meant torefer to bicyclic and polycyclic hydrocarbon rings such as, for example,bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane, etc. The term“heterocycloalkyl” refers to a cycloalkyl group that contain from one tofive heteroatoms selected from N, O, and S, wherein the nitrogen andsulfur atoms are optionally oxidized, and the nitrogen atom(s) areoptionally quaternized. The heterocycloalkyl may be a monocyclic, abicyclic or a polycylic ring system. Non limiting examples ofheterocycloalkyl groups include pyrrolidine, imidazolidine,pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin,dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine,thiomorpholine, thiomorpholine-S-oxide, thiomorpholine-S,S-oxide,piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone,tetrahydrofuran, tetrhydrothiophene, quinuclidine, and the like. Aheterocycloalkyl group can be attached to the remainder of the moleculethrough a ring carbon or a heteroatom.

The term “alkylene” by itself or as part of another substituent means adivalent radical derived from an alkane, as exemplified by—CH₂CH₂CH₂CH₂—. Typically, an alkyl (or alkylene) group will have from 1to 24 carbon atoms, with those groups having 10 or fewer carbon atomsbeing preferred in the present invention. A “lower alkyl” or “loweralkylene” is a shorter chain alkyl or alkylene group, generally havingfour or fewer carbon atoms. Similarly, “alkenylene” and “alkynylene”refer to the unsaturated forms of “alkylene” having double or triplebonds, respectively.

As used herein, a wavy line, “

”, that intersects a single, double or triple bond in any chemicalstructure depicted herein, represent the point attachment of the single,double, or triple bond to the remainder of the molecule. A bondrepresented by

is meant to depict an optional double bond. As such, the symbol refersto either a single bond or a double bond.

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. Additionally, for dialkylaminogroups, the alkyl portions can be the same or different and can also becombined to form a 3-7 membered ring with the nitrogen atom to whicheach is attached. Accordingly, a group represented as dialkylamino or—NR^(a)R^(b) is meant to include piperidinyl, pyrrolidinyl, morpholinyl,azetidinyl and the like.

The term “di-(C₁₋₄ alkyl)amino-C₁₋₄ alkyl” refers to an amino groupbearing two C₁₋₄ alkyl groups that can be the same or different (e.g.,methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl andtert-butyl) and which is attached to the remainder of the moleculethrough a C₁₋₄ alkyl group (a one to four carbon alkylene linkinggroup). Examples of di-(C₁₋₄ alkyl)amino-C₁₋₄ alkyl groups includedimethylaminomethyl, 2-(ethyl(methyl)amino)ethyl,3-(dimethylamino)butyl, and the like.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “and acid isosteres” means, unless otherwise stated, a groupwhich can replace a carboxylic acid, having an acidic functionality andsteric and electronic characteristics that provide a level of activity(or other compound characteristic such as solubility) similar to acarboxylic acid. Representative acid isosteres include, hydroxamicacids, sulfonic acids, sulfinic acids, sulfonamides, acyl-sulfonamides,phosphonic acids, phosphinic acids, phosphoric acids, tetrazole, andoxo-oxadiazoles.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of aryl groups include phenyl, naphthyl and biphenyl, whilenon-limiting examples of heteroaryl groups include pyridyl, pyridazinyl,pyrazinyl, pyrimindinyl, triazinyl, quinolinyl, quinoxalinyl,quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl,benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl,isobenzofuryl, isoindolyl, indolizinyl, benzotriazinyl, thienopyridinyl,thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines,benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl,isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl,triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl,thiazolyl, furyl, thienyl and the like. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

The term “arylalkyl” is meant to include those radicals in which an arylgroup is attached to an alkyl group (e.g., benzyl, phenethyl, and thelike). Similarly, the term “heteroaryl-alkyl” is meant to include thoseradicals in which a heteroaryl group is attached to an alkyl group(e.g., pyridylmethyl, thiazolylethyl, and the like).

The above terms (e.g., “alkyl,” “aryl” and “heteroaryl”), in someembodiments, will recite both substituted and unsubstituted forms of theindicated radical. Preferred substituents for each type of radical areprovided below.

Substituents for the alkyl radicals (including those groups oftenreferred to as alkylene, alkenyl, alkynyl and cycloalkyl) can be avariety of groups selected from: -halogen, —OR′, —NR′R″, —SR′,—SiR′R″R′″, —OC(O)R′, —C(O)R′, —CO₂R′, —CONR′R″, —OC(O)NR′R″,—NR″C(O)R′, —NR′—C(O)NR″R′″, —NR″C(O)₂R′, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR'S(O)₂R″, —CN and—NO₂ in a number ranging from zero to (2 m′+1), where m′ is the totalnumber of carbon atoms in such radical. R′, R″ and R′″ eachindependently refer to hydrogen, unsubstituted C₁₋₈ alkyl, unsubstitutedaryl, aryl substituted with 1-3 halogens, unsubstituted C₁₋₈ alkyl, C₁₋₈alkoxy or C₁₋₈ thioalkoxy groups, or unsubstituted aryl-C₁₋₄ alkylgroups. When R′ and R″ are attached to the same nitrogen atom, they canbe combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or7-membered ring. For example, —NR′R″ is meant to include 1-pyrrolidinyland 4-morpholinyl.

Similarly, substituents for the aryl and heteroaryl groups are variedand are generally selected from: -halogen, —OR′, —OC(O)R′, —NR′R″, —SR′,—R′, —CN, —NO₂, —CO₂R′, —CONR′R″, —C(O)R′, —OC(O)NR′R″, —NR″C(O)R′,—NR″C(O)₂R′, —NR′—C(O)NR″R′″, —NH—C(NH₂)═NH, —NR′C(NH₂)═NH,—NH—C(NH₂)═NR′, —S(O)R′, —S(O)₂R′, —S(O)₂NR′R″, —NR′S(O)₂R″, —N₃,perfluoro(C₁-C₄)alkoxy, and perfluoro(C₁-C₄)alkyl, in a number rangingfrom zero to the total number of open valences on the aromatic ringsystem; and where R′, R″ and R′″ are independently selected fromhydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₃₋₆ cycloalkyl, C₂₋₈ alkenyl andC₂₋₈ alkynyl. Other suitable substituents include each of the above arylsubstituents attached to a ring atom by an alkylene tether of from 1-4carbon atoms.

Two of the substituents on adjacent atoms of the aryl or heteroaryl ringmay optionally be replaced with a substituent of the formula-T-C(O)—(CH₂)_(q)—U—, wherein T and U are independently —NH—, —O—, —CH₂—or a single bond, and q is an integer of from 0 to 2. Alternatively, twoof the substituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula-A-(CH₂)_(r)—B—, wherein A and B are independently —CH₂—, —O—, —NH—,—S—, —S(O)—, —S(O)₂—, —S(O)₂NR′— or a single bond, and r is an integerof from 1 to 3. One of the single bonds of the new ring so formed mayoptionally be replaced with a double bond. Alternatively, two of thesubstituents on adjacent atoms of the aryl or heteroaryl ring mayoptionally be replaced with a substituent of the formula—(CH₂)_(s)—X—(CH₂)_(t)—, where s and t are independently integers offrom 0 to 3, and X is —O—, —NR′—, —S—, —S(O)—, —S(O)₂—, or —S(O)₂NR′—.The substituent R′ in —NR′— and —S(O)₂NR′— is selected from hydrogen orunsubstituted C₁₋₆ alkyl.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of salts derived frompharmaceutically-acceptable inorganic bases include aluminum, ammonium,calcium, copper, ferric, ferrous, lithium, magnesium, manganic,manganous, potassium, sodium, zinc and the like. Salts derived frompharmaceutically-acceptable organic bases include salts of primary,secondary and tertiary amines, including substituted amines, cyclicamines, naturally-occuring amines and the like, such as arginine,betaine, caffeine, choline, N,N′-dibenzylethylenediamine, diethylamine,2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine,ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine,glucosamine, histidine, hydrabamine, isopropylamine, lysine,methylglucamine, morpholine, piperazine, piperidine, polyamine resins,procaine, purines, theobromine, triethylamine, trimethylamine,tripropylamine, tromethamine and the like. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers, regioisomers and individual isomers (e.g., separateenantiomers) are all intended to be encompassed within the scope of thepresent invention. The compounds of the present invention may alsocontain unnatural proportions of atomic isotopes at one or more of theatoms that constitute such compounds. Unnatural proportions of anisotope may be defined as ranging from the amount found in nature to anamount consisting of 100% of the atom in question. For example, thecompounds may incorporate radioactive isotopes, such as for exampletritium (³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C), or non-radioactiveisotopes, such as deuterium (²H) or carbon-13 (¹³C). Such isotopicvariations can provide additional utilities to those described elsewherewith this application. For instance, isotopic variants of the compoundsof the invention may find additional utility, including but not limitedto, as diagnostic and/or imaging reagents, or as cytotoxic/radiotoxictherapeutic agents. Additionally, isotopic variants of the compounds ofthe invention can have altered pharmacokinetic and pharmacodynamiccharacteristics which can contribute to enhanced safety, tolerability orefficacy during treatment. All isotopic variations of the compounds ofthe present invention, whether radioactive or not, are intended to beencompassed within the scope of the present invention.

The “subject” is defined herein to include animals such as mammals,including, but not limited to, primates (e.g., humans), cows, sheep,goats, horses, dogs, cats, rabbits, rats, mice and the like. Inpreferred embodiments, the subject is a human.

As used herein, the phrase “CCR(4)-mediated condition or disease” andrelated phrases and terms refer to a condition or disease characterizedby inappropriate, e.g., less than or greater than normal, CCR(4)functional activity. Inappropriate CCR(4) functional activity mightarise as the result of CCR(4) expression in cells which normally do notexpress CCR(4), increased CCR(4) expression (leading to, e.g.,inflammatory and immunoregulatory disorders and diseases) or decreasedCCR(4) expression. Inappropriate CCR(4) functional activity might alsoarise as the result of TARC and/or MDC secretion by cells which normallydo not secrete TARC and/or MDC, increased TARC and/or MDC expression(leading to, e.g., inflammatory and immunoregulatory disorders anddiseases) or decreased T ARC and/or MDC expression. A CCR(4)-mediatedcondition or disease may be completely or partially mediated byinappropriate CCR(4) functional activity. However, a CCR(4)-mediatedcondition or disease is one in which modulation of CCR(4) results insome effect on the underlying condition or disease (e.g., a CCR(4)antagonist results in some improvement in patient well-being in at leastsome patients).

The term “therapeutically effective amount” means the amount of thesubject compound that will elicit the biological or medical response ofa tissue, system, animal or human that is being sought by theresearcher, veterinarian, medical doctor or other clinician.

II. General

Compounds of the present invention can modulate CCR(4) function and areuseful in the treatment of various inflammatory and immunoregulatorydisorders and diseases.

III. Embodiments of the Invention

A. Compounds

In one aspect, the present invention provides compounds having FormulaI:

and any pharmaceutically acceptable salts thereof, wherein

-   -   R¹ is selected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl, C₁₋₈        hydroxyalkyl, C₃₋₈ cycloalkyl, halogen, —CN, —SO₂Me and        —C(O)NH₂;    -   each R² is selected from C₁₋₈ alkyl, C₁₋₈ haloalkyl, halogen,        —CN and C₁₋₈ alkoxy; or two R² groups attached to adjacent        carbon atoms are optionally connected to form a 5 or 6 member        ring (aliphatic or aromatic, heterocycle or not) which is        optionally substituted with additional R² groups;    -   R³ is selected from hydrogen, methyl and C₁₋₄ haloalkyl;    -   R⁴ is selected from hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and        C₁₋₈ hydroxyalkyl;    -   each of the subscripts n is independently an integer from 0 to        3;    -   B is a bond or C(O);    -   Q is a selected from C, CH, N, O, S, S(O), and SO₂;    -   W, X, Y, and Z are independently selected from C, CH and N, but        Q and W are not both N;    -   R⁵ and R⁶ are absent or are independently selected from H, —OH,        C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₄ alkoxy-C₁₋₄ alkyl,        —C(O)NR^(a)R^(b), C₁₋₈ alkylene-C(O)NR^(a)R^(b), —NH—C₁₋₄        alkylene-C(O)NR^(a)R^(b), —C(O)—C₁₋₄ alkylene-NR^(a)R^(b), —CO₂H        and acid isosteres, C₁₋₈ alkylene-CO₂H and acid isosteres,        —N(R^(a))C(O)NR^(a)R^(b), C₁₋₈ alkylene-N(R^(a))C(O)NR^(a)R^(b),        —NR^(a)R^(b), —R^(a)R^(b), C₁₋₈ alkylene-NR^(a)R^(b), C₁₋₈        alkoxy, —C(O)OR^(a), C₁₋₈ alkylene-C(O)OR^(a), —CN, —C(O)R^(a),        —SO₂R^(a) and —N(R^(a))C(O)R^(b);    -   wherein        -   each R^(a) and R^(b) are independently selected from            hydrogen, C₁₋₈ alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl, and            C₁₋₈ alkoxy; and    -   R⁷ is absent or is selected from H, C₁₋₈ alkyl and C₁₋₈        haloalkyl.

In one group of embodiments, the compounds provided herein are thosewherein X and Y are not both N. In another group of embodiments, R³ isH, and each R² is independently selected from C₁₋₈ alkyl, C₁₋₈haloalkyl, halogen and —CN.

In another group of embodiments, the compounds provided herein have theformula (Ia):

-   -   wherein each of R¹, R², R⁴, R⁵, R⁶, R⁷, X, Y, Z, W, Q, B and the        subscripts n, are as described for formula I. In selected        embodiments, X is C or CH.

In another group of embodiments, the compounds provided herein have theformula (Ib):

wherein each R² is selected from C₁₋₈ alkyl, C₁₋₈ haloalkyl, halogen and—CN, and each of R¹, R⁴, R⁵, R⁶, R⁷, X, Z, W, Q, B and the subscripts n,are as described for formula I.

In another group of embodiments, the compounds provided herein have theformula (Ic):

wherein each R² is selected from C₁₋₈ alkyl, C₁₋₈ haloalkyl, halogen and—CN; the subscript n is 0 or 1; and each of R¹, R⁴, R⁶, R⁷, X, Y, Z, andQ, are as described for formula I. In selected embodiments, n is 1, andR⁴ is hydrogen or methyl.

In another group of embodiments, the compounds provided herein have theformula (Id):

wherein each R² is a member selected from C₁₋₈ alkyl, C₁₋₈ haloalkyl,halogen and —CN; the subscript n is 0 or 1, and each of R¹, R⁴, R⁶, R⁷,X, Z, and Q, are as described for formula I. In selected embodiments, nis 1, and R⁴ is hydrogen or methyl.

In another group of embodiments, the compounds provided herein have theformula (Ie):

wherein each R² is selected from C₁₋₈ alkyl, C_(1-g) haloalkyl, halogenand —CN; the subscript n is 0 or 1; and each of R¹, R⁴, R⁶, R⁷, Y, Z,and Q, are as described for formula I. In selected embodiments, n is 1,and R⁴ is hydrogen or methyl.

In another group of embodiments, the compounds provided herein have theformula (If):

wherein each R² is selected from C₁₋₈ alkyl, C₁₋₈ haloalkyl, halogen and—CN; and each of R¹, R⁴, R⁶, R⁷, and Q, are as described for formula I.In selected embodiments, R⁴ is hydrogen or methyl.

In still other embodiments, compounds are provided having formulae (I),(Ia), and (Ib), including specific embodiments provided above, wherein Bis C(O). Still further, compounds are provided wherein the ring having Zas a ring vertex is selected from pyrrolidine and piperidine. Inselected embodiments, compounds are provided wherein the ring having Zas a ring vertex is selected from pyrrolidin-2-yl and piperidin-2-yl,and at least one of R⁵, R⁶ and R⁷ is other than hydrogen

In yet other embodiments, compounds are provided having formulae (I),(Ia), and (Ib), including specific embodiments provided above, wherein Bis a bond. In related embodiments, B is a bond and the ring having Z asa ring vertex is selected from pyrrolidine, piperidine and cyclohexane.In specific embodiments, B is a bond and the ring having Z as a ringvertex is selected from pyrrolidin-1-yl, pyrrolidin-2-yl,piperidin-1-yl, piperidin-2-yl, piperidin-3-yl and cyclohexane. In stillother embodiments, B is a bond and the ring having Z as a ring vertex isselected from the group consisting of pyrrolidin-1-yl, pyrrolidin-2-yl,piperidin-1-yl, piperidin-2-yl, piperidin-3-yl and cyclohexane; and atleast one of R⁵, R⁶ and R⁷ is other than hydrogen.

In one group of embodiments, Z is CH or N.

In a selected group of embodiments, compounds are provided having thestructures:

B. Compositions

In addition to the compounds provided above, compositions for modulatingCCR(4) activity in humans and animals will typically contain apharmaceutical carrier or diluent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The pharmaceutical compositions for the administration of the compoundsof this invention may conveniently be presented in unit dosage form andmay be prepared by any of the methods well known in the art of pharmacyand drug delivery. All methods include the step of bringing the activeingredient into association with the carrier which constitutes one ormore accessory ingredients. In general, the pharmaceutical compositionsare prepared by uniformly and intimately bringing the active ingredientinto association with a liquid carrier or a finely divided solid carrieror both, and then, if necessary, shaping the product into the desiredformulation. In the pharmaceutical composition the active objectcompound is included in an amount sufficient to produce the desiredeffect upon the process or condition of diseases.

The pharmaceutical compositions containing the active ingredient may bein a form suitable for oral use, for example, as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules,emulsions and self emulsifications as described in U.S. PatentApplication 2002-0012680, hard or soft capsules, syrups, elixirs,solutions, buccal patch, oral gel, chewing gum, chewable tablets,effervescent powder and effervescent tablets. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents selected from the group consisting ofsweetening agents, flavoring agents, coloring agents, antioxidants andpreserving agents in order to provide pharmaceutically elegant andpalatable preparations. Tablets contain the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for the manufacture of tablets. These excipients may be forexample, inert diluents, such as cellulose, silicon dioxide, aluminumoxide, calcium carbonate, sodium carbonate, glucose, mannitol, sorbitol,lactose, calcium phosphate or sodium phosphate; granulating anddisintegrating agents, for example, corn starch, or alginic acid;binding agents, for example PVP, cellulose, PEG, starch, gelatin oracacia, and lubricating agents, for example magnesium stearate, stearicacid or talc. The tablets may be uncoated or they may be coated,enterically or otherwise, by known techniques to delay disintegrationand absorption in the gastrointestinal tract and thereby provide asustained action over a longer period. For example, a time delaymaterial such as glyceryl monostearate or glyceryl distearate may beemployed. They may also be coated by the techniques described in theU.S. Pat. Nos. 4,256,108; 4,166,452; and U.S. Pat. No. 4,265,874 to formosmotic therapeutic tablets for control release.

Formulations for oral use may also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example peanut oil, liquid paraffin, or olive oil.Additionally, emulsions can be prepared with a non-water miscibleingredient such as oils and stabilized with surfactants such asmono-diglycerides, PEG esters and the like.

Aqueous suspensions contain the active materials in admixture withexcipients suitable for the manufacture of aqueous suspensions. Suchexcipients are suspending agents, for example sodiumcarboxymethylcellulose, methylcellulose, hydroxy-propylmethylcellulose,sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia;dispersing or wetting agents may be a naturally-occurring phosphatide,for example lecithin, or condensation products of an alkylene oxide withfatty acids, for example polyoxy-ethylene stearate, or condensationproducts of ethylene oxide with long chain aliphatic alcohols, forexample heptadecaethyleneoxycetanol, or condensation products ofethylene oxide with partial esters derived from fatty acids and ahexitol such as polyoxyethylene sorbitol monooleate, or condensationproducts of ethylene oxide with partial esters derived from fatty acidsand hexitol anhydrides, for example polyethylene sorbitan monooleate.The aqueous suspensions may also contain one or more preservatives, forexample ethyl, or n-propyl, p-hydroxybenzoate, one or more coloringagents, one or more flavoring agents, and one or more sweetening agents,such as sucrose or saccharin.

Oily suspensions may be formulated by suspending the active ingredientin a vegetable oil, for example arachis oil, olive oil, sesame oil orcoconut oil, or in a mineral oil such as liquid paraffin. The oilysuspensions may contain a thickening agent, for example beeswax, hardparaffin or cetyl alcohol. Sweetening agents such as those set forthabove, and flavoring agents may be added to provide a palatable oralpreparation. These compositions may be preserved by the addition of ananti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueoussuspension by the addition of water provide the active ingredient inadmixture with a dispersing or wetting agent, suspending agent and oneor more preservatives. Suitable dispersing or wetting agents andsuspending agents are exemplified by those already mentioned above.Additional excipients, for example sweetening, flavoring and coloringagents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, forexample olive oil or arachis oil, or a mineral oil, for example liquidparaffin or mixtures of these. Suitable emulsifying agents may benaturally-occurring gums, for example gum acacia or gum tragacanth,naturally-occurring phosphatides, for example soy bean, lecithin, andesters or partial esters derived from fatty acids and hexitolanhydrides, for example sorbitan monooleate, and condensation productsof the said partial esters with ethylene oxide, for examplepolyoxyethylene sorbitan monooleate. The emulsions may also containsweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, for exampleglycerol, propylene glycol, sorbitol or sucrose. Such formulations mayalso contain a demulcent, a preservative and flavoring and coloringagents. Oral solutions can be prepared in combination with, for example,cyclodextrin, PEG and surfactants.

The pharmaceutical compositions may be in the form of a sterileinjectable aqueous or oleagenous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally-acceptable diluent orsolvent, for example as a solution in 1,3-butane diol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium. For this purpose any bland fixed oil may be employedincluding synthetic mono- or diglycerides. In addition, fatty acids suchas oleic acid find use in the preparation of injectables.

The compounds of the present invention may also be administered in theform of suppositories for rectal administration of the drug. Thesecompositions can be prepared by mixing the drug with a suitablenon-irritating excipient which is solid at ordinary temperatures butliquid at the rectal temperature and will therefore melt in the rectumto release the drug. Such materials include cocoa butter andpolyethylene glycols. Additionally, the compounds can be administeredvia ocular delivery by means of solutions or ointments. Still further,transdermal delivery of the subject compounds can be accomplished bymeans of iontophoretic patches and the like. For topical use, creams,ointments, jellies, solutions or suspensions, etc., containing thecompounds of the present invention are employed. As used herein, topicalapplication is also meant to include the use of mouth washes andgargles.

The compounds of this invention may also be coupled a carrier that is asuitable polymers as targetable drug carriers. Such polymers can includepolyvinylpyrrolidone, pyran copolymer, polyhydroxy-propyl-methacrylamide-phenol, polyhydroxyethyl-aspartamide-phenol, orpolyethyleneoxide-polylysine substituted with palmitoyl residues.Furthermore, the compounds of the invention may be coupled to a carrierthat is a class of biodegradable polymers useful in achieving controlledrelease of a drug, for example polylactic acid, polyglycolic acid,copolymers of polylactic and polyglycolic acid, polyepsiloncaprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals,polydihydropyrans, polycyanoacrylates and cross linked or amphipathicblock copolymers of hydrogels. Polymers and semipermeable polymermatrices may be formed into shaped articles, such as valves, stents,tubing, prostheses and the like.

C. Methods of Use

In another aspect, the present disclosure provides methods of treatingor preventing a CCR(4)-mediated condition or disease by administering toa subject having such a condition or disease, a therapeuticallyeffective amount of any compound of Formula I. Preferred compounds foruse in the present methods are those compounds provided herein aspreferred embodiments, as well as compounds specifically set forth inthe Examples below, in the attached Figures; and provided with specificstructures herein.

Diseases and conditions associated with inflammation, infection andcancer can be treated or prevented with the present compounds andcompositions. In one group of embodiments, diseases or conditions,including chronic diseases, of humans or other species can be treatedwith inhibitors of CCR(4) function. These diseases or conditionsinclude: (1) allergic diseases such as systemic anaphylaxis orhypersensitivity responses, drug allergies, insect sting allergies andfood allergies, (2) inflammatory bowel diseases, such as Crohn'sdisease, ulcerative colitis, ileitis and enteritis, (3) vaginitis, (4)psoriasis and inflammatory dermatoses such as dermatitis, eczema, atopicdermatitis, allergic contact dermatitis, dermatomyositis, lichen planus,bullous pemphigoid, urticaria and pruritus, (5) vasculitis, (6)spondyloarthropathies, (7) scleroderma, (8) asthma and respiratoryallergic diseases such as allergic asthma, exercise-induced asthma,allergic rhinitis, hypersensitivity lung diseases and the like, (9)autoimmune diseases, such as arthritis (including rheumatoid andpsoriatic), multiple sclerosis, systemic lupus erythematosus, type Idiabetes, glomerulonephritis, and the like, (10) graft rejection(including allograft rejection and graft-v-host disease), and (11)leukemias, lymphomas, and other blood borne cancers including cutaneousT cell lymphoma, mycosis fungoides, acute lymphoblastic leukemias andthe like, and (12) other diseases in which undesired inflammatoryresponses are to be inhibited, such as atherosclerosis, myositis,neurodegenerative diseases (e.g., Alzheimer's disease), encephalitis,meningitis, hepatitis, nephritis, sepsis, sarcoidosis, allergicconjunctivitis, otitis, chronic obstructive pulmonary disease,sinusitis, Behcet's syndrome and gout.

In another group of embodiments, diseases or conditions can be treatedwith agonists of CCR(4) function. Examples of diseases to be treatedwith CCR(4) agonists include cancers, diseases in which angiogenesis orneovascularization play a role (neoplastic diseases, retinopathy andmacular degeneration), infectious diseases (viral infections, e.g., HIVinfection, and bacterial infections) and immunosuppressive diseases suchas organ transplant conditions and skin transplant conditions. The term“organ transplant conditions” is meant to include bone marrow transplantconditions and solid organ (e.g., kidney, liver, lung, heart, pancreasor combination thereof) transplant conditions.

Preferably, the present methods are directed to the treatment ofdiseases or conditions selected from allergic diseases (including skinallergies and allergic airway disorders), atopic allergic conditionsincluding atopic dermatitis, psoriasis, cancer (including solid tumorsand metastatic disease) and asthma. Depending on the disease to betreated and the subject's condition, the compounds of the presentinvention may be administered by oral, parenteral (e.g., intramuscular,intraperitoneal, intravenous, ICV, intracisternal injection or infusion,subcutaneous injection, or implant), inhalation, nasal, vaginal, rectal,sublingual, or topical routes of administration and may be formulated,alone or together, in suitable dosage unit formulations containingconventional non-toxic pharmaceutically acceptable carriers, adjuvantsand vehicles appropriate for each route of administration. The presentinvention also contemplates administration of the compounds of thepresent invention in a depot formulation.

Those of skill in the art will understand that agents that modulateCCR(4) activity can be combined in treatment regimens with othertherapeutic agents and/or with chemotherapeutic agents or radiation. Insome cases, the amount of chemotherapeutic agent or radiation is anamount which would be sub-therapeutic if provided without combinationwith a composition of the invention. Those of skill in the art willappreciate that “combinations” can involve combinations in treatments(i.e., two or more drugs can be administered as a mixture, or at leastconcurrently or at least introduced into a subject at different timesbut such that both are in the bloodstream of a subject at the sametime). Additionally, compositions of the current invention may beadministered prior to or subsequent to a second therapeutic regimen, forinstance prior to or subsequent to a dose of chemotherapy orirradiation.

In the treatment or prevention of conditions which require chemokinereceptor modulation an appropriate dosage level will generally be about0.001 to 100 mg per kg patient body weight per day which can beadministered in single or multiple doses. Preferably, the dosage levelwill be about 0.01 to about 25 mg/kg per day; more preferably about 0.05to about 10 mg/kg per day. A suitable dosage level may be about 0.01 to25 mg/kg per day, about 0.05 to 10 mg/kg per day, or about 0.1 to 5mg/kg per day. Within this range the dosage may be 0.005 to 0.05, 0.05to 0.5 or 0.5 to 5.0 mg/kg per day. For oral administration, thecompositions are preferably provided in the form of tablets containing1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 5.0,10.0, 15.0. 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0,400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of theactive ingredient for the symptomatic adjustment of the dosage to thepatient to be treated. The compounds may be administered on a regimen of1 to 4 times per day, preferably once or twice per day.

It will be understood, however, that the specific dose level andfrequency of dosage for any particular patient may be varied and willdepend upon a variety of factors including the activity of the specificcompound employed, the metabolic stability and length of action of thatcompound, the age, body weight, hereditary characteristics, generalhealth, sex and diet of the subject, as well as the mode and time ofadministration, rate of excretion, drug combination, and the severity ofthe particular condition for the subject undergoing therapy.

In one group of embodiments, the compounds and compositions describedherein can be combined with other compounds and compositions havingrelated utilities to prevent and treat cancer and diseases or conditionsassociated with CCR(4) signaling. Such other drugs may be administered,by a route and in an amount commonly used therefor, contemporaneously orsequentially with a compound or composition of the present invention.When a compound or composition of the present invention is usedcontemporaneously with one or more other drugs, a pharmaceuticalcomposition containing such other drugs in addition to the compound orcomposition of the present invention is preferred. Accordingly, thepharmaceutical compositions of the present invention include those thatalso contain one or more other active ingredients or therapeutic agents,in addition to a compound or composition of the present invention.Examples of other therapeutic agents that may be combined with acompound or composition of the present invention, either administeredseparately or in the same pharmaceutical compositions, include, but arenot limited to: cisplatin, paclitaxel, methotrexate, cyclophosphamide,ifosfamide, chlorambucil, carmustine, carboplatin, vincristine,vinblastine, thiotepa, lomustine, semustine, 5-fluorouracil,corticosteroids, calcineurin inhibitors, NSAIDs, inhibitors of5-lipoxygenase, and cytarabine. The weight ratio of the compound of thepresent invention to the second active ingredient may be varied and willdepend upon the effective dose of each ingredient. Generally, aneffective dose of each will be used. Thus, for example, when a compoundof the present invention is combined with a second anticancer agent, theweight ratio of the compound of the present invention to the secondagent will generally range from about 1000:1 to about 1:1000, preferablyabout 200:1 to about 1:200. Combinations of a compound of the presentinvention and other active ingredients will generally also be within theaforementioned range, but in each case, an effective dose of each activeingredient should be used.

Methods of Treating Inflammation

Still further, the compounds and compositions of the present inventionare useful for the treatment of inflammation, and can be combined withother compounds and compositions having therapeutic utilities that mayrequire treatment either before, after or simultaneously with thetreatment of cancer or inflammation with the present compounds.Accordingly, combination methods and compositions are also a componentof the present invention to prevent and treat the condition or diseaseof interest, such as inflammatory or autoimmune disorders, conditionsand diseases, including psoriasis, dermatomyositis, inflammatory boweldisease, rheumatoid arthritis, osteoarthritis, psoriatic arthritis,polyarticular arthritis, multiple sclerosis, allergic diseases, atopicdermatitis and asthma, and those pathologies noted above.

For example, in the treatment or prevention of inflammation orautimmunity or for example arthritis associated bone loss, the presentcompounds and compositions may be used in conjunction with ananti-inflammatory or analgesic agent such as an opiate agonist, alipoxygenase inhibitor, such as an inhibitor of 5-lipoxygenase, acyclooxygenase inhibitor, such as a cyclooxygenase-2 inhibitor, aninterleukin inhibitor, such as an interleukin-1 inhibitor, an NMDAantagonist, an inhibitor of nitric oxide or an inhibitor of thesynthesis of nitric oxide, a non steroidal anti-inflammatory agent, or acytokine-suppressing anti-inflammatory agent, for example with acompound such as acetaminophen, aspirin, codeine, fentanyl, ibuprofen,indomethacin, ketorolac, morphine, naproxen, phenacetin, piroxicam, asteroidal analgesic, sufentanyl, sunlindac, tenidap, and the like.Similarly, the instant compounds and compositions may be administeredwith an analgesic listed above; a potentiator such as caffeine, an H2antagonist (e.g., ranitidine), simethicone, aluminum or magnesiumhydroxide; a decongestant such as phenylephrine, phenylpropanolamine,pseudoephedrine, oxymetazoline, ephinephrine, naphazoline,xylometazoline, propylhexedrine, or levo desoxy ephedrine; anantitussive such as codeine, hydrocodone, caramiphen, carbetapentane, ordextromethorphan; a diuretic; and a sedating or non sedatingantihistamine.

As noted, compounds and compositions of the present invention may beused in combination with other drugs that are used in the treatment,prevention, suppression or amelioration of the diseases or conditionsfor which compounds and compositions of the present invention areuseful. Such other drugs may be administered, by a route and in anamount commonly used therefor, contemporaneously or sequentially with acompound or composition of the present invention. When a compound orcomposition of the present invention is used contemporaneously with oneor more other drugs, a pharmaceutical composition containing such otherdrugs in addition to the compound or composition of the presentinvention is preferred. Accordingly, the pharmaceutical compositions ofthe present invention include those that also contain one or more otheractive ingredients or therapeutic agents, in addition to a compound orcomposition of the present invention. Examples of other therapeuticagents that may be combined with a compound or composition of thepresent invention, either administered separately or in the samepharmaceutical compositions, include, but are not limited to: (a) VLA-4antagonists, (b) corticosteroids, such as beclomethasone,methylprednisolone, betamethasone, prednisone, prenisolone,dexamethasone, fluticasone, hydrocortisone, budesonide, triamcinolone,salmeterol, salmeterol, salbutamol, formeterol; (c) immunosuppressantssuch as cyclosporine (cyclosporine A, Sandimmune®, Neoral®), tacrolirnus(FK-506, Prograf®), rapamycin (sirolimus, Rapamune®) and other FK-506type immunosuppressants, and rnycophenolate, e.g., mycophenolate mofetil(CellCept®); (d) antihistamines (H1-histamine antagonists) such asbromopheniramine, chlorpheniramine, dexchloipheniramine, triprolidine,clemastine, diphenhydramine, diphenylpyraline, tripelennamine,hydroxyzine, methdilazine, promethazine, trimeprazine, azatadine,cyproheptadine, antazoline, pheniramine pyrilamine, astemizole,terfenadine, loratadine, cetirizine, fexofenadine,descarboethoxyloratadine, and the like; (e) non steroidal antiasthmatics (e.g., terbutaline, metaproterenol, fenoterol, isoetharine,albuterol, bitolterol and pirbuterol), theophylline, cromolyn sodium,atropine, ipratropium bromide, leukotriene antagonists (e.g.,zafmlukast, montelukast, pranlukast, iralukast, pobilukast andSKB-106,203), leukotriene biosynthesis inhibitors (zileuton, BAY-1005);(f) non steroidal anti-inflammatory agents (NSAIDs) such as propionicacid derivatives (e.g., alminoprofen, benoxaprofen, bucloxic acid,carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, ibuprofen,indoprofen, ketoprofen, rniroprofen, naproxen, oxaprozin, pirprofen,pranoprofen, suprofen, tiaprofenic acid and tioxaprofen), acetic acidderivatives (e.g., indomethacin, acemetacin, alclofenac, clidanac,diclofenac, fenclofenac, fenclozic acid, fentiazac, furofenac, ibufenac,isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidometacin andzomepirac), fenamic acid derivatives (e.g., flufenamic acid,meclofenamic acid, mefenamic acid, niflumic acid and tolfenamic acid),biphenylcarboxylic acid derivatives (e.g., diflunisal and flufenisal),oxicams (e.g., isoxicam, piroxicam, sudoxicam and tenoxican),salicylates (e.g., acetyl salicylic acid and sulfasalazine) and thepyrazolones (e.g., apazone, bezpiperylon, feprazone, mofebutazone,oxyphenbutazone and phenylbutazone); (g) cyclooxygenase-2 (COX-2)inhibitors such as celecoxib (Celebrex®) and rofecoxib (Vioxx®); (h)inhibitors of phosphodiesterase type IV (PDE IV); (i) gold compoundssuch as auranofin and aurothioglucose, (j) TNF-alpha modulators such asetanercept (Enbrel®), (k) antibody therapies such as orthoclone (OKT3),daclizumab (Zenapax®), basiliximab (Simulect®), B cell modulators suchas rituximab (Rituxan®), and infliximab (Remicade®), (1) otherantagonists of the chemokine receptors, especially CCR1, CCR5, CXCR2,CXCR3, CCR2, CCR3, CCR(4), CCR7, CCR9, CX₃CR1 and CXCR6; (m) lubricantsor emollients such as petrolatum and lanolin, (n) keratolytic agents(e.g., tazarotene), (o) vitamin D₃ derivatives, e.g., calcipotriene orcalcipotriol (Dovonex®), (p) PUVA, (q) anthralin (Drithrocreme®), (r)etretinate (Tegison®) and isotretinoin and (s) multiple sclerosistherapeutic agents such as interferon (3-1f3 (Betaseron®), interferon(P3-la (Avonex®), azathioprine (Imurek®, Imuran®), glatiramer acetate(Capoxone®), a glucocorticoid (e.g., prednisolone) and cyclophosphamide(t) DMARDS such as methotrexate (u) T cell costimulatory modulators suchas abatacept (Orencia®), (v) other compounds such as 5-aminosalicylicacid and prodrugs thereof; hydroxychloroquine; D-penicillamine;antimetabolites such as azathioprine, 6-mercaptopurine and methotrexate;DNA synthesis inhibitors such as hydroxyurea and microtubule disrupterssuch as colchicine. The weight ratio of the compound of the presentinvention to the second active ingredient may be varied and will dependupon the effective dose of each ingredient. Generally, an effective doseof each will be used. Thus, for example, when a compound of the presentinvention is combined with an NSAID the weight ratio of the compound ofthe present invention to the NSAID will generally range from about1000:1 to about 1:1000, preferably about 200:1 to about 1:200.Combinations of a compound of the present invention and other activeingredients will generally also be within the aforementioned range, butin each case, an effective dose of each active ingredient should beused.

IV. Examples

The following examples are offered to illustrate, but not to limit theclaimed invention.

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). 1H-NMR spectra wererecorded on a Varian Mercury 400 MHz NMR spectrometer. Significant peaksare provided relative to TMS and are tabulated in the order:multiplicity (s, singlet; d, doublet; t, triplet; q, quartet; m,multiplet) and number of protons. Mass spectrometry results are reportedas the ratio of mass over charge, followed by the relative abundance ofeach ion (in parenthesis). In the examples, a single m/e value isreported for the M+H (or, as noted, M−H) ion containing the most commonatomic isotopes. Isotope patterns correspond to the expected formula inall cases. Electrospray ionization (ESI) mass spectrometry analysis wasconducted on a Hewlett-Packard MSD electrospray mass spectrometer usingthe HP 1100 HPLC for sample delivery. Normally the analyte was dissolvedin methanol at 0.1 mg/mL and 1 microlitre was infused with the deliverysolvent into the mass spectrometer, which scanned from 100 to 1500daltons. All compounds could be analyzed in the positive ESI mode, usingacetonitrile/water with 1% formic acid as the delivery solvent. Thecompounds provided below could also be analyzed in the negative ESImode, using 2 mM NH₄OAc in acetonitrile/water as delivery system.

The following abbreviations are used in the Examples and throughout thedescription of the invention: rt, room temperature; HPLC, high pressureliquid chromatography; TFA, trifluoroacetic acid; LC-MSD, liquidchromatograph/mass selective detector; LC-MS, liquid chromatograph/massspectrometer; Pd₂dba₃, tris(dibenzylideneacetone) dipalladium; THF,tetrahydrofuran; DMF, dimethylformamide or N,N-dimethylformamide; DCM,dichloromethane; DMSO, dimethyl sulfoxide; TLC, thin-layerchromatography; KHMDS, potassium hexamethyldisilazane; ES, electrospray;sat., saturated.

Compounds within the scope of this invention can be synthesized asdescribed below, using a variety of reactions known to the skilledartisan. One skilled in the art will also recognize that alternativemethods may be employed to synthesize the target compounds of thisinvention, and that the approaches described within the body of thisdocument are not exhaustive, but do provide broadly applicable andpractical routes to compounds of interest.

Certain molecules claimed in this patent can exist in differentenantiomeric and diastereomeric forms and all such variants of thesecompounds are claimed.

The detailed description of the experimental procedures used tosynthesize key compounds in this text lead to molecules that aredescribed by the physical data identifying them as well as by thestructural depictions associated with them.

Those skilled in the art will also recognize that during standard workup procedures in organic chemistry, acids and bases are frequently used.Salts of the parent compounds are sometimes produced, if they possessthe necessary intrinsic acidity or basicity, during the experimentalprocedures described within this patent.

Examples Preparation of Compounds

Those skilled in the art will recognize that there are a variety ofmethods available to synthesize molecules represented in the claims. Ingeneral, useful methods for synthesizing compounds represented in theclaims consist of five parts, which may be done in any order: formationof the chiral amine, formation of the bicycle system, coupling betweenthe chiral amine and the bicycle, installation of substituents at Q orW, and installation and/or modification of functional groups on thevarious substituents.

Several methods for the preparation of the compounds are illustrated inFIGS. 1-8 (see eq. 1-18). Equations 1-2 demonstrate methods of formingof the chiral amine. Equations 3-10 demonstrate some methods ofpreparing of the bicycles via amide bond formation or amine formation.Coupling of the chiral amine and the bicycle viz metal-mediated couplingare shown in equations 11-14. Equations 15-18 demonstrate methods tointroduce substitution at Q or W then results in the compounds of theinvention.

Examples Example 1: Resolution of (1R)-1-(2,4-dichlorophenyl)ethanamine

(S)-Mandelic acid (40.2 g, 264.5 mmol) was added to a solution of 3:2isopropyl alcohol (iPrOH) and ethanol (EtOH) (500 mL) at roomtemperature, and the suspension was heated at 60° C. until a clearsolution formed. Racemic 2,4-dichloro-α-methyl benzylamine (50 g, 264.5mmol) was added to the hot solution, which was then cooled to 30° C.over 2 h and stirred at this temperature for 24 h. The colorlesscrystals were collected by filtration and washed with acetone (70 mL).The resulting salt (37.3 g, ˜90% ee, determined by Mosher's method, J.Am. Chem. Soc., 1973, 95, 512.) was suspended in 3:2 iPrOH/EtOH (400 mL)at room temperature and the mixture was heated at 60° C. to give a clearsolution. The solution was then cooled to room temperature and stirredfor 24 h. The colorless crystals were filtered off, and washed withacetone (40 mL) to give the desired salt (32.0 g, >96% ee, determined byto Mosher's method). To a portion of the salt (12.0 g) indichloromethane (CH₂Cl₂) (100 mL) was added aqueous 4 N sodium hydroxidesolution (30 mL). The reaction mixture was stirred for 1 h at roomtemperature, and extracted with dichloromethane (2×100 mL), dried withanhydrous sodium sulfate (Na₂SO₄), filtered, and concentrated in vacuoto afford (1R)-1-(2,4-dichlorophenyl)ethanamine as a colorless liquid(7.5 g, 39.5 mmol, 40%).

Example 2: Synthesis of2-chloro-N-((1R)-1-(2,4-dichlorophenyl)ethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(see FIG. 5)

a) To a stirred solution of 3-bromo-4-chlorophenol (25.0 g, 120.5 mmol)in dichloromethane at 0° C. was added imidazole (9.0 g, 132.5 mmol) andt-butyldimethylchlorosilane (TBSCl) (19.0 g, 126.5 mmol). The solutionwas stirred at 0° C. for 30 min, then at room temperature for 14 h. Thereaction mixture was diluted with deionized water, extracted withdichloromethane (2×200 mL). The combined organic layers were dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude product waspurified by flash chromatography (SiO₂, 0-10% ethyl acetate in hexanes)to afford (3-bromo-4-chlorophenoxy)-t-butyldimethylsilane as colorlessoil (37.0 g, 115.0 mmol, 98%). MS: (ES) m/z calculated for C₁₂H₁₉BrClOSi[M+H]⁺ 321.0, found 321.

b) To a mixture of (3-bromo-4-chlorophenoxy)-t-butyldimethylsilane (15.0g, 46.8 mmol) and (1R)-1-(2,4-dichlorophenyl)ethanamine (9.7 g, 51.5mmol) in anhydrous toluene (250 mL) were added palladium(II) acetate(Pd(OAc)₂) (0.42 g, 1.9 mmol),2,2′-bis(diphenylphosphino)-1,1′-binaphthyl (BINAP) (1.74 g, 2.80 mmol),and sodium t-butoxide (NaO-tBu) (6.3 g, 65.52 mmol). The resultingmixture was heated at 110° C. under nitrogen for 3 h. After cooling toroom temperature, the suspension was filtered through a plug of Celite,and washed with ethyl acetate (EtOAc) (100 mL). The filtrate wasconcentrated in vacuo. The resulting residue was diluted with deionizedwater, and extracted with ethyl acetate (500 mL). The organic layer waswashed with brine, dried (Na₂SO₄), and concentrated in vacuo. The cruderesidue was purified by flash chromatography (SiO₂, 0-15% ethyl acetatein hexanes) to afford(R)-5-(t-butyldimethylsilyloxy)-2-chloro-N-(1-(2,4-dichlorophenyl)ethyl)anilineas a viscous oil (18.5 g, 42.9 mmol, 92%). ¹H NMR (400 MHz, CDCl₃) δ7.42 (d, J=1.6 Hz, 1H), 7.32 (d, J=8.8 Hz, 1H), 7.19 (dd, J=1.6, 8.4 Hz,1H), 7.08 (d, J=8.4 Hz, 1H), 6.13 (dd, J=2.2, 8.4 Hz, 1H), 5.71 (d,J=2.4 Hz, 1H), 4.87-4.84 (m, 1H), 4.67 (d, J=4.8 Hz, 1H), 1.58 (d, J=6.8Hz, 3H), 0.98 (s, 9H), 0.02 (s, 3H), 0.01 (s, 3H).

c) To a stirred solution of(R)-5-(t-butyldimethylsilyloxy)-2-chloro-N-(1-(2,4-dichlorophenyl)ethyl)aniline(18.5 g, 43.0 mmol) in anhydrous tetrahydrofuran (THF) (100 mL) at 0° C.was added a solution of tetrabutylammonium fluoride (TBAF) (1 M solutionin THF, 43.0 mL, 43.0 mmol). The reaction mixture was stirred at 0° C.for 30 min, and then warmed to room temperature and stirred for anadditional 4 h. The reaction mixture was diluted with deionized waterand extracted with ethyl acetate (2×50 mL). The combined organic layerswere washed with deionized water, brine, dried (Na₂SO₄), filtered, andconcentrated in vacuo to give(R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenol as a brown solid(15.3 g). The solid compound was carried to the next step withoutfurther purification.

d) To a stirred solution of the(R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenol (18.0 g, 56.9mmol) in dichloromethane (150 mL) at 0° C. was added triethylamine(Et₃N) (9.6 g, 91.0 mmol) and trifluoromethanesulfonic anhydride (TFA)(17.6 g, 62.6 mmol). The reaction mixture was stirred at 0° C. for 2 h,then warm to room temperature and stirred for 10 min. The solution wasdiluted with deionized water, extracted with dichloromethane (2×50 mL).The combined organic layers were washed with brine, dried (Na₂SO₄),filtered, and concentrated in vacuo. The crude product was purified byflash chromatography (SiO₂, 0-25% ethyl acetate in hexanes) to afford(R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyltrifluoromethanesulfonate as viscous oil (17.8 g, 39.7 mmol, 72%). ¹HNMR (400 MHz, CDCl₃) δ 7.40 (d, J=2.0 Hz, 1H), 7.25-7.23 (m, 2H), 7.16(dd, J=1.6, 8.4 Hz, 1H), 6.49 (dd, J=2.8, 8.8 Hz, 1H), 6.05 (d, J=3.2,Hz, 1H), 4.91-4.89 (m, 1H), 4.88-4.83 (m, 1H), 1.58 (d, J=6.8 Hz, 3H).

e) A mixture of (R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyltrifluoromethanesulfonate (6.0 g, 13.3 mmol), bis(pinacolato)diboron(4.08 g, 15.9 mmol),[1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II)(Pd(dppf)Cl₂) (0.39 g, 0.53 mmol), and potassium acetate (3.93 g, 39.9mmol) in p-dioxane (100 mL) was purged with nitrogen for 5 min, and thenheated at 95° C. for 4 h. After cooling to room temperature, the mixturewas filtered and washed with EtOAc (50 mL). The filtrate wasconcentrated in vacuo. The resulting crude mixture was diluted withethyl acetate (200 mL), washed with deionized water, brine, dried(Na₂SO₄), and concentrated in vacuo. The resulting crude was purified byflash chromatography (SiO₂, 0-15% ethyl acetate in hexanes) to affordthe title compound as a white solid (4.5 g, 10.5 mmol, 79%). ¹H NMR (400MHz, CDCl₃) δ 7.35 (d, J=6.0 Hz, 1H), 7.27 (d, J=8.4 Hz, 1H), 7.22 (d,J=5.2 Hz, 1H), 7.12 (dd, J=2.0, 8.4 Hz, 1H), 6.98 (dd, J=0.8, 7.6 Hz,1H), 6.77 (s, 1H), 5.03-4.98 (m, 1H), 4.66 (d, J=6.4 Hz, 1H), 1.52 (d,J=6.8 Hz, 3H), 1.27 (s, 6H), 1.25 (s, 6H); MS: (ES) m/z calculated forC₂₀H₂₄BCl₃NO₂ [M+H]⁺ 426.1, found 426.

Example 3: Synthesis of2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)-5-(1-((S)-3-piperidyl)-3,6-dihydro-2H-pyridin-4-yl)aniline(see FIG. 6)

a) Methyl iodide (7.3 mL, 117.8 mmol) was slowly added to a solution ofthe N-ethylpiperidone (13.4 mL, 98.2 mmol) in acetone (100 mL) at roomtemperature. The reaction mixture was stirred for 5 h and a solid wasformed. The colorless solid was collected by filtration, washed withacetone (30 mL), and dried under vacuum to give the quaternary salt (25g, 92.9 mmol, 95%).

b) N-Ethyl-N-methyl-4-oxo-piperidinium iodide (8.0 g, 29.7 mmol)dissolved in deionized water (30 mL) was added to a refluxing solutionof (S)-1-boc-3-aminopiperidine (5 g, 25 mmol) and potassium carbonate(K₂CO₃) (3.5 g, 25 mmol) in ethanol (125 mL) at 80° C. The heating wascontinued for 4 h and excess solvent was removed in vacuo. The aqueouslayer was extracted with ethyl acetate (3×60 mL), and the combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The residue was purified by flash chromatography (SiO₂, 1-5% methanol indichloromethane) to give the desired compound (5.8 g, 20.5 mmol, 82%).MS: (ES) m/z calculated for C₁₅H₂₇N₂O₃ [M+H]⁺ 283.2, found 283.

c) To a solution of 1-((3S)-3-piperidyl)piperidin-4-one (3.3 g, 11.68mmol) in anhydrous THF (20 mL) at −78° C. under nitrogen was slowlyadded a solution of lithium diisopropylamide (LDA) (2 M solution in THF,7 mL, 14 mmol). After the addition is complete, the reaction mixture waswarmed to −20° C. over 30 min. The solution was re-cooled to −78° C.,and a solution of N-phenyl-bis(trifluoromethanesulfonimide) (5.4 g, 15.2mmol) dissolved in anhydrous THF (20 mL) was added dropwise. Thereaction mixture was warmed to room temperature over 6 h, and quenchedwith saturated aqueous ammonium chloride (NH₄Cl) solution. The aqueouslayer was extracted with ethyl acetate (2×50 mL). The combined organiclayers were dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresidue was purified flash chromatography (SiO₂, 0-20% ethyl acetate indichloromethane) to give the pure compound (2.5 g, 6.0 mmol, 52%). MS:(ES) m/z calculated for C₁₆H₂₆F₃N₂O₅S [M+H]⁺ 415.4, found 415.

d) To a solution (S)-t-butyl3-(4-(trifluoromethylsulfonyloxy)-5,6-dihydropyridin-1(2H)-yl)piperidine-1-carboxylate(2.0 g, 4.8 mmol) and(R)-2-chloro-N-(1-(2,4-dichlorophenyl)ethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(prepared from example 2, 2.3 g, 5.3 mmol) in dimethoxyethane (12 mL)was added tetrakis(triphenylphosphine)palladium(0) (Pd(PPh₃)₄) (0.28 g,0.24 mmol) and 2 M aqueous potassium carbonate solution (6 mL, 12 mmol).The mixture was purged with nitrogen for 5 min, and heated at 85° C.under nitrogen for 5 h. After cooling to room temperature, the reactionmixture was diluted with ethyl acetate. The organic layer was washedwith brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. Theresidue was purified by flash chromatography (SiO₂, 0-5% methanol indichloromethane) to give the pure compound (1.4 g, 2.5 mmol, 51%). MS:(ES) m/z calculated for C₁₆H₂₆F₃N₂O₅S [M+H]⁺ 564.2, found 564.

e) Trifluoroacetic acid (1.5 mL) was added to a solution of (S)-t-butyl3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)piperidine-1-carboxylate(0.83 g, 1.5 mmol) in dichloromethane (3 mL) at room temperature, andthe solution was stirred for 1 h. Excess solvent was removed in vacuo,and the residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was used without further purification (0.65 g, 1.4mmol, 96%). MS: (ES) m/z calculated for C₂₄H₂₉ClN₃ [M+H]⁺ 464.1, found464.

Example 4: Synthesis of2-((S)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)piperidin-1-yl)aceticacid (see FIG. 7)

A mixture of the crude2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)-5-(1-((S)-3-piperidyl)-3,6-dihydro-2H-pyridin-4-yl)aniline(0.10 g, 0.21 mmol), 2-bromomethyl acetate (0.036 g, 0.24 mmol), andpotassium carbonate (0.058 g, 0.42 mmol) in 1-methyl-2-pyrrolidinone(NMP) (0.8 mL) was stirred under nitrogen for 12 h. The reaction mixturewas diluted with ethyl acetate, washed with deionized water, brine,dried (Na₂SO₄), filtered, and concentrated in vacuo. The residue waspurified by flash chromatography (SiO₂, 0-5% methanol indichloromethane) to give the desired compound (0.040 g, 0.075 mmol,35%). MS: (ES) m/z calculated for C₂₇H₃₃Cl₃N₃O₂ [M+H]⁺ 536.2, found 536.The resulting ester was dissolved in methanol (0.14 mL) and THF (0.6mL), and aqueous 1 N sodium hydroxide solution (0.14 mL, 0.14 mmol) wasadded. The reaction mixture was heated at 50° C. for 1 h, and cooled toroom temperature. The solution was diluted with deionized water (1 mL),and the crude was purified by reverse phase HPLC (C18 column,acetonitrile-H₂O with 0.1% TFA as eluent) to give a light yellow solidas TFA salt (0.037 g, 0.052 mmol, 66%). ¹H NMR (400 MHz, CD₃OD) δ 7.45(d, J=2.8 Hz, 1H), 7.41 (d, J=8.4 Hz, 1H), 7.25-7.21 (m, 2H), 6.72 (dd,J=2.2, 8.4 Hz, 1H), 6.31 (d, J=2.2 Hz, 1H), 5.91 (bs, 1H), 5.01 (q,J=7.0 Hz, 1H), 4.00-3.90 (m, 2H), 3.70-3.44 (m, 6H), 3.23-3.18 (m, 1H),3.00 (bs, 1H), 2.90-2.82 (m, 1H), 2.70 (bs, 2H), 2.10-1.90 (m, 5H),1.84-1.74 (m, 1H), 1.58 (d, J=6.6 Hz, 3H); MS: (ES) m/z calculated forC₂₆H₃₁Cl₃N₃O₂ [M+H]⁺ 522.2, found 522.

Example 5: Synthesis of3-((S)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)piperidin-1-yl)propanoicacid (see FIG. 8)

The titled compound was prepared as illustrated in example 4 to give alight yellow solid as TFA salt. ¹H NMR (400 MHz, CD₃OD) δ 7.44 (d, J=2.8Hz, 1H), 7.41 (d, J=8.4 Hz, 1H), 7.25-7.21 (m, 2H), 6.71 (dd, J=2.2, 8.0Hz, 1H), 6.29 (d, J=2.2 Hz, 1H), 5.91 (bs, 1H), 5.00 (q, J=6.6 Hz, 1H),4.00-3.70 (m, 5H), 3.60-3.40 (m, 6H), 3.25-3.21 (m, 1H), 3.00-2.92 (m,1H), 2.88 (t, J=6.9 Hz, 2H), 2.78-2.64 (m, 2H), 2.30-2.10 (m, 2H),1.96-1.80 (m, 2H), 1.58 (d, J=6.6 Hz, 3H); MS: (ES) m/z calculated forC₂₇H₃₃Cl₃N₃O₂ [M+H]⁺ 536.1, found 536.

Example 6: Synthesis of5-((S)-1,3′-bipiperidin-4-yl)-2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)aniline(see FIG. 9)

A mixture of the crude2-chloro-N-((1R)-1-(2,4-dichlorophenyl)ethyl)-5-(1-((3S)-3-piperidyl)-3,6-dihydro-2H-pyridin-4-yl)aniline(0.30 g, 0.63 mmol), platinum(IV) oxide (PtO₂) (5% by weight, 0.045 g)in ethanol (3 mL) containing concentrated hydrochloric acid (conc. HCl)(0.20 mL, 2.20 mmol) in a Paar shaker flask was hydrogenated at 40 psifor 45 min. The reaction mixture was diluted with ethanol, and filteredthrough the Celite. The filtrate was concentrated in vacuo, and theresulting residue was diluted with dichloromethane (25 mL). The organiclayer was washed with saturated aqueous sodium bicarbonate solution,dried (Na₂SO₄), filtered, and concentrated in vacuo. The residue wasused without further purification (0.27 g, 0.058 mmol). MS: (ES) m/zcalculated for C₂₄H₃₁Cl₃N₃[M+H]⁺ 466.2, found 466.

Example 7: Synthesis of2-((S)-4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-1,3′-bipiperidin-1′-yl)aceticacid (see FIG. 10)

The titled compound was prepared as illustrated in example 4 to give awhite solid as TFA salt. ¹H NMR (400 MHz, CD₃OD) δ 7.46 (d, J=2.2 Hz,1H), 7.40 (d, J=8.0 Hz, 1H), 7.26-7.24 (m, 1H), 7.20 (d, J=8.0 Hz, 1H),6.52 (dd, J=1.9, 8.1 Hz, 1H), 6.15 (d, J=1.9 Hz, 1H), 5.01 (q, J=6.6 Hz,1H), 3.74-3.39 (m, 6H), 3.20-3.06 (m, 5H), 2.87-2.80 (m, 1H), 2.72-2.64(m, 1H), 2.10-1.78 (m, 9H), 1.57 (d, J=6.6 Hz, 3H); MS: (ES) m/zcalculated for C₂₆H₃₃Cl₃N₃O₂ [M+H]⁺ 524.2, found 524.

Example 8: Synthesis of3-((S)-4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-1,3′-bipiperidin-1′-yl)propanoicacid (see FIG. 11)

The titled compound was prepared as illustrated in example 4 to give awhite solid as TFA salt. ¹H NMR (400 MHz, CD₃OD) δ 7.44 (d, J=2.2 Hz,1H), 7.40 (d, J=8.4 Hz, 1H), 7.26-7.23 (m, 1H), 7.20 (d, J=8.1 Hz, 1H),6.51 (dd, J=1.9, 8.1 Hz, 1H), 6.15 (d, J=2.2 Hz, 1H), 4.98 (q, J=6.6 Hz,1H), 3.91-3.88 (m, 1H), 3.74-3.66 (m, 1H), 3.59-3.56 (m, 3H), 3.50-3.42(m, 2H), 3.30-3.18 (m, 5H), 3.00-2.92 (m, 1H), 2.89 (t, J=7.0 Hz, 2H),2.72-2.64 (m, 1H), 2.28-2.15 (m, 2H), 2.02-1.81 (m, 6H), 1.57 (d, J=6.6Hz, 3H); MS: (ES) m/z calculated for C₂₇H₃₅C₁₃N₃O₂ [M+H]⁺ 538.2, found538.

Example 9: Synthesis of(1R,3R)-3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-1-yl)cyclohexanecarboxamide(see FIG. 12)

a) A mixture of (R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyltrifluoromethanesulfonate (prepared from Example 2 step d, 5.0 g, 11.4mmol), (N-t-butoxycarbonyl-1,2,3,6-tetrahydropyridin-4-yl)boronic acidpincol ester (5.8 g, 14.8 mmol), Pd(dppf)Cl₂ (0.5 g, 0.068 mmol), andpotassium phosphate tribasic (K₃PO₄) (6.0 g, 34.2 mmol) in p-dioxane(100 mL) was purged with nitrogen for 5 min, and then heated at 100° C.for 1 h. After cooling to room temperature, the mixture was filtered andwashed with ethyl acetate (50 mL). The filtrate was concentrated invacuo. The resulting crude mixture was diluted with ethyl acetate (200mL), washed with deionized water, brine, dried (Na₂SO₄), andconcentrated in vacuo. The resulting crude was purified by flashchromatography (SiO₂, 0-20% methanol in dichloromethane) to afford thedesired product (4.8 g, 9.5 mmol, 84%).

b) Trifluoroacetic acid (2.0 mL) was added to a solution of (R)-t-butyl4-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridine-1(2H)-carboxylate(4.8 g, 9.5 mmol) in dichloromethane (6 mL) at room temperature, and thesolution was stirred for 1 h. Excess solvent was removed in vacuo, andthe residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was used without further purification (3.2 g, 8.4mmol, 89%).

c) To a stirred solution of the crude(R)-2-chloro-N-(1-(2,4-dichlorophenyl)ethyl)-5-(1,2,3,6-tetrahydropyridin-4-yl)aniline(0.65 g, 0.17 mmol) and 3-oxocyclohexanecarboxylic acid (0.036 g, 0.26mmol) in dichloromethane (4 mL) was added sodium triacetoxyborohydride(NaBH(OAc)₃) (0.054 g, 0.25 mmol). The reaction mixture was stirred atroom temperature for 18 h. The reaction mixture was quenched withaqueous saturated sodium bicarbonate and extracted with dichloromethane.The organic layer was dried (Na₂SO₄), filtered, and concentrated invacuo. The resulting crude was purified by flash chromatography (SiO₂,10-40% ethyl acetate in hexanes) to afford3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)cyclohexanecarboxylicacid as the desired product (0.064 g, 0.13 mmol, 74%).

d) To a stirred solution of the3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)cyclohexanecarboxylic acid (0.070 g, 0.14 mmol) andN,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)uroniumhexafluorophosphate (HATU) (0.063 g, 0.165 mmol) in anhydrousN,N-dimethylforamide (DMF) (2.5 mL) was added a solution of ammonia (0.5M in p-dioxane, 2.7 mL, 1.4 mmol) and iPr₂NEt (0.045 g, 0.34 mmol). Thereaction was heated at 45° C. for 3 h. After cooling to roomtemperature, the reaction mixture was quenched with deionized water,extracted with diethyl ether. The organic layer was dried (Na₂SO₄),filtered, and concentrated in vacuo. The crude product was purified byflash chromatography (SiO₂, 0-15% methanol in dichloromethane) to affordthe desired product (0.042 g, 0.083 mmol, 60%).

e) A mixture of the3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)-5,6-dihydropyridin-1(2H)-yl)cyclohexanecarboxamide(0.154 g, 0.20 mmol), palladium on carbon (Pd/C) (10% by weight, 0.05 g)in ethanol (7 mL) containing concentrated hydrochloric acid (4 drops) ina Paar shaker flask was hydrogenated at 25 psi for 1 h. The reactionmixture was diluted with ethanol, and filtered through Celite. Thefiltrate was concentrated in vacuo, and the resulting residue waspurified by flash chromatography (SiO₂, 5% triethylamine in ethylacetate) to separate the desired isomer as a white solid (0.038 g, 0.075mmol, 25%). ¹H NMR (400 MHz, CD₃OD) δ 7.45-7.41 (m, 2H), 7.26 (dd,J=2.0, 8.4 Hz, 1H), 7.21 (d, J=8.0 Hz, 1H), 6.57 (d, J=8.0 Hz, 1H), 6.21(s, 1H), 4.99 (dd, J=6.8, 13.2 Hz, 1H), 3.76-3.71 (m, 1H), 3.53-3.52 (m,3H), 3.20-3.12 (m, 2H), 2.99-2.93 (m, 1H), 2.71-2.65 (m, 1H), 2.47 (d,J=11.6 Hz, 1H), 2.35 (d, J=13.2 Hz, 1H), 2.18-2.10 (m, 2H), 1.99-1.82(m, 6H), 1.68-1.74 (m, 1H), 1.64-1.51 (m, 6H); MS: (ES) m/z calculatedfor C₂₆H₃₃Cl₃N₃O [M+H]⁺ 508.2, found 508.4.

Example 10: Synthesis of(R)-(1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)(morpholino)methanone(see FIG. 13)

a) A mixture of (R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyltrifluoromethanesulfonate (Example 2 step d, 1.5 g, 3.4 mmol), ethyl4-piperidinecarboxylate (1.18 g, 8.4 mmol), tBuDavePhos (0.12 g, 0.34mmol), and cesium carbonate (Cs₂CO₃) (2.7 g, 8.4 mmol) in triethylamine(12 mL) was purged with nitrogen for 5 min. Addition of Pd₂(dba)₃ (0.15g, 0.14 mmol) was followed, and the mixture was purged with nitrogen for1 min. The reaction was then heated at 100° C. for 18 h. After coolingto room temperature, the mixture was filtered and washed with EtOAc (50mL). The filtrate was concentrated in vacuo. The resulting crude mixturewas purified by flash chromatography (SiO₂, 0-15% ethyl acetate inhexanes) to afford the coupled product as a viscous oil (0.82, 1.82mmol, 52%).

b) To a stirred solution of(R)-ethyl1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidine-4-carboxylate(3.1 g, 6.8 mmol) in 2:1 THF/EtOH (12 mL) was added a solution of sodiumhydroxide (1.4 g, 34.1 mmol) dissolved in deionized water (4 mL). Thereaction mixture was stirred at room temperature for 4 h, and themixture was adjusted to pH-7 with aqueous 6 N hydrochloric acid. Theaqueous layer was extracted with ethyl acetate (2×50 mL). The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was purified by flash chromatography (SiO₂, 0-25%methanol in dichloromethane) to give(R)-1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidine-4-carboxylicacid (2.4 g, 5.6 mmol, 82%).

c) To a stirred solution of the(R)-1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidine-4-carboxylicacid (0.0.05 g, 0.12 mmol) and N-methylmorpholine (0.01 g, 0.12 mmol) inDMF (1.5 mL) was added HATU (0.049 g, 0.13 mmol) and iPr₂NEt (0.75 g,5.8 mmol). The reaction mixture was stirred at room temperature for 2 h,and quenched with deionized water. The aqueous layer was extracted withdiethyl ether (2×10 mL). The organic layer was dried (Na₂SO₄), filtered,and concentrated in vacuo. The crude product purified by reverse phaseHPLC (C18 column, acetonitrile-H₂O with 0.1% TFA as eluent) to give awhite solid (0.025 g, 0.050 mmol, 43%). ¹H NMR (400 MHz, CDCl₃) δ7.36-7.33 (m, 1H), 7.17 (dd, J=1.6, 8.4 Hz, 1H), 7.06 (d, J=8.8 Hz, 1H),6.18 (dd, J=2.8, 8.4 Hz, 1H), 5.78 (d, J=2.4 Hz, 1H), 4.90 (ddd, J=6.4,6.4, 12.4 Hz, 1H), 4.65 (d, J=5.2 Hz, 1H), 3.68-3.63 (m, 5H), 3.50-3.47(m, 3H), 2.64-2.48 (m, 3H), 1.92-1.80 (m, 5H), 1.71-1.67 (m, 2H), 1.53(d, J=6.4 Hz, 3H); MS: (ES) m/z calculated for C₂₄H₂₉Cl₃N30₂ [M+H]+496.1, found 496.3.

Example 11: Synthesis of(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)((S)-3-methoxypyrrolidin-1-yl)methanone(see FIG. 14)

The titled compound was prepared as illustrated in Example 10 c using(3S)-methoxypyrrolidine as the coupling partner to give a white solid.¹H NMR (400 MHz, CDCl₃) δ 7.36-7.32 (m, 2H), 7.16 (dd, J=2.2, 8.4 Hz,1H), 7.04 (d, J=8.6 Hz, 1H), 6.16 (dd, J=2.5, 8.8 Hz, 1H), 5.78 (d,J=2.6 Hz, 1H), 4.87 (q, J=6.6 Hz, 1H), 4.62 (d, J=5.2 Hz, 1H), 4.06-3.92(m, 1H), 3.70-3.42 (m, 6H), 3.32 (s, 3H). 2.64-2.32 (m, 3H), 2.18-1.70(m, 6H), 1.52 (d, J=6.6 Hz, 3H); MS: (ES) m/z calculated forC₂₅H₃₁Cl₃N₃O₂ [M+H]⁺ 510.1, found 510.

Example 12: Synthesis of1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidine-4-carbonyl)pyrrolidine-3-carbonitrile(see

FIG. 15)

The titled compound was prepared as illustrated in Example 10 c using3-pyrrolidinecarbonitrile as the coupling partner to give a white solid.¹H NMR (400 MHz, CDCl₃) δ 7.37-7.33 (m, 2H), 7.17 (dd, J=2.0, 8.0 Hz,1H), 7.06 (d, J=8.4 Hz, 1H), 6.18 (dd, J=2.8, 8.4 Hz, 1H), 5.78 (d,J=2.8 Hz, 1H), 4.90 (ddd, J=6.4, 6.4, 12.4 Hz, 1H), 4.64 (d, J=5.2 Hz,1H), 3.86-3.70 (m, 2H), 3.67-3.58 (m, 1H), 3.51-3.45 (m, 2H), 3.23-3.12(m, 1H), 2.96 (s, 1H), 2.89 (s, 1H), 2.61-2.53 (m, 2H), 2.39-2.19 (m,3H), 1.87-1.72 (m, 3H), 1.53 (d, J=6.4 Hz, 3H); MS: (ES) m/z calculatedfor C₂₅H₂₈Cl₃N₄O [M+H]⁺ 505.1, found 505.2.

Example 13: Synthesis of(4-(3-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazin-1-yl)((R)-piperidin-2-yl)methanone(see FIG. 16)

a) To a stirred solution of 5-bromo-2-fluorophenol (3.8 g, 20.0 mmol) indichloromethane (50 mL) at 0° C. was added iPr₂NEt (1.5 g, 11.5 mmol)and t-butyldimethylchlorosilane (TBSCl) (3.6 g, 24.1 mmol). The mixturewas stirred at 0° C. for 30 min, then at room temperature for 14 h. Thereaction mixture was diluted with deionized water and extracted withdichloromethane (2×50 mL). The combined organic layers were dried(Na₂SO₄), filtered, and concentrated in vacuo. The crude was usedwithout further purification (5.9 g, 19.4 mmol, 97%).

b) A mixture of (5-bromo-2-chlorophenoxy)-t-butyldimethylsilane (1.23 g,4.04 mmol), N-t-butyloxycarbonylpiperazine (1.58 g, 10.1 mmol),tBuDavePhos (0.14 g, 0.4 mmol), and Cs₂CO₃ (3.5 g, 11.7 mmol) in 2:1p-dioxane/triethylamine (12 mL) was purged with nitrogen for 10 min.Addition of Pd₂(dba)₃ (0.15 g, 0.16 mmol) was followed, and the mixturewas purged with nitrogen for 1 min. The reaction was then heated at 100°C. for 4.5 h. After cooling to room temperature, the mixture wasfiltered and washed with EtOAc (50 mL). The filtrate was concentrated invacuo. The resulting crude mixture was purified by flash chromatography(SiO₂, 5-30% ethyl acetate in hexanes) to yieldt-butyl-4-(3-t-butyldimethylsilyloxy)-4-fluorophenyl)piperazine-1-carboxylate(1.5 g, 3.73 mmol, 92%).

c) To a stirred solution oft-butyl-4-(3-t-butyldimethylsilyloxy)-4-fluorophenyl)piperazine-1-carboxylate(2.5 g, 6.2 mmol) in anhydrous tetrahydrofuran (THF) (20 mL) added asolution of tetrabutylammonium fluoride (TBAF) (1 M solution in THF, 6.2mL, 6.2 mmol), and the reaction mixture was stirred at room temperaturefor 1 h. The solution was diluted with deionized water and extractedwith ethyl acetate (2×20 mL). The combined organic layers were washedwith deionized water, brine, dried (Na₂SO₄), filtered, and concentratedin vacuo to give t-butyl4-(4-fluoro-3-hydroxyphenyl)piperazine-1-carboxylate (1.5 g, 5.1 mmol,86%). The solid compound was carried to the next step with out furtherpurification.

d) To a stirred solution of the t-butyl4-(4-fluoro-3-hydroxyphenyl)piperazine-1-carboxylate (1.35 g, 4.56 mmol)in dichloromethane (15 mL) at 0° C. was added triethylamine (0.75 g,7.48 mmol) and TFA (1.58 g, 5.62 mmol). The reaction mixture was stirredat 0° C. for 15 min, then warm to room temperature and stirred for 30min. The solution was diluted with deionized water, extracted withdichloromethane (2×10 mL). The combined organic layers were washed withbrine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudeproduct was purified by flash chromatography (SiO₂, 5-25% ethyl acetatein hexanes) to afford t-butyl4-(4-fluoro-3-(trifluoromethylsulfonyloxy)phenyl)piperazine-1-carboxylate(1.53 g, 3.57 mmol, 76%).

e) To a mixture of t-butyl4-(4-fluoro-3-(trifluoromethylsulfonyloxy)phenyl)piperazine-1-carboxylate(0.50 g, 1.17 mmol) and (1R)-1-(2,4-dichlorophenyl)ethanamine (0.58 g,3.05 mmol), tBuDavePhos (0.025 g, 0.070 mmol), and Cs₂CO₃ (0.80 g, 2.44mmol) in triethylamine (3 mL) was purged with nitrogen for 3 min.Addition of Pd₂(dba)₃ (0.034 g, 0.037 mmol) was followed, and themixture was purged with nitrogen for 1 min. The resulting mixture washeated at 100° C. under nitrogen for 4 h. After cooling to roomtemperature, the suspension was filtered through a plug of Celite, andwashed with ethyl acetate (25 mL). The filtrate was concentrated invacuo and the resulting residue purified by flash chromatography (SiO₂,10-50% ethyl acetate in hexanes) to afford (R)-t-butyl4-(3-(1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazine-1-carboxylate(0.36 g, 42.9 mmol, 62%).

f) Trifluoroacetic acid (1.5 mL) was added to a solution of (R)-t-butyl4-(3-(1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazine-1-carboxylate(0.52 g, 1.11 mmol) in dichloromethane (5 mL) at room temperature, andthe solution was stirred for 1 h. Excess solvent was removed in vacuo,and the residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was used without further purification (0.38 g, 1.4mmol, 94%).

g) To a stirred solution of the(R)—N-(1-(2,4-dichlorophenyl)ethyl)-2-fluoro-5-(piperazin-1-yl)aniline(0.085 g, 0.23 mmol) and (R)-1-(t-butoxycarbonyl)piperdine-2-carboxylicacid (0.053 g, 0.23 mmol) in DMF (1.5 mL) was added HATU (0.11 g, 0.28mmol) and iPr₂NEt (0.075 g, 0.58 mmol). The reaction mixture was stirredat room temperature for 2 h, and quenched with deionized water. Theaqueous layer was extracted with diethyl ether (2×10 mL). The organiclayer was dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudeproduct was purified by flash chromatography (SiO₂, 10-70% ethyl acetatein hexanes) to afford the desired product (0.080 g, 0.14 mmol, 65%).

h) Trifluoroacetic acid (1.5 mL) was added to a solution (R)-t-butyl2-(4-(3-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazine-1-carbonyl)piperidine-1-carboxylate(0.080 g, 0.14 mmol) in dichloromethane (5 mL) at room temperature, andthe solution was stirred for 1 h. Excess solvent was removed in vacuo,and the residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuoto give the title compound as a white solid (0.042 g, 0.088 mmol, 64%).¹H NMR (400 MHz, DMSO-d₆) δ 9.25 (s, 1H), 8.60-8.55 (m, 1H), 7.56 (dd,J=8.4, 8.4 Hz, 2H), 7.38 (dd, J=2.4, 8.4 Hz, 1H), 6.92 (dd, J=9.2, 11.2Hz, 1H), 6.26-6.09 (m, 3H), 4.83 (dd, J=6.4, 13.6 Hz, 1H), 4.37 (t,J=11.2 Hz, 1H), 3.76-3.48 (m, 4H), 3.21 (d, J=11.2 Hz, 1H), 3.09-3.05(m, 2H), 2.90-2.84 (m, 2H), 1.95 (d, J=13.6 Hz, 1H), 1.73-1.60 (m, 3H),1.45 (d, J=6.8 Hz, 3H); MS: (ES) m/z calculated for C₂₄H₃₀Cl₂FN₄O [M+H]⁺479.2, found 479.4.

Example 14: Synthesis of(4-(3-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-fluorophenyl)piperazin-1-yl)((R)-pyrrolidin-2-yl)methanone(see FIG. 17)

The titled compound was prepared as illustrated in Example 13 g-h using(R)-1-t-butoxycarbonylpyrrolidine-2-carboxylic acid as the couplingpartner to give a white solid as the title compound. ¹H NMR (400 MHz,DMSO-d₆) δ 10.0 (s, 1H), 8.47-8.44 (m, 2H), 7.56 (dd, J=2.0, 7.6 Hz,1H), 7.38 (dd, J=2.0, 8.4 Hz, 1H), 6.92 (dd, J=10.4, 10.4 Hz, 1H), 6.26(bs, 1H), 6.09 (bs, 1H), 4.83 (q, J=6.6 Hz, 1H), 4.59 (ddd, J=6.8, 6.8,13.2 Hz, 1H), 3.67-3.65 (m, 4H), 3.24-3.13 (m, 2H), 3.01 (s, 4H),2.38-2.33 (m, 1H), 1.93-1.76 (m, 3H), 1.49 (d, J=6.8 Hz, 3H); MS: (ES)m/z calculated for C₂₃H₂₈Cl₂FN₄O [M+H]+ 465.2, found 465.3.

Example 15: Synthesis of3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexanecarboxamide(see FIG. 18)

a) To a stirred solution of(R)-2-chloro-N-(1-(2,4-dichlorophenyl)ethyl)-5-(piperazin-1-yl)aniline(prepared as illustrated in example 13 using 5-bromo-2-chlorophenol asstarting material, 0.20 g, 0.54 mmol) and 3-oxocyclohexanecarboxylicacid (0.15 g, 1.1 mmol) in dichloromethane (4 mL) was added NaBH(OAc)₃(0.153 g, 1.1 mmol). The reaction mixture was stirred at roomtemperature for 18 h, and quenched with diluted with aqueous saturatedsodium bicarbonate. The aqueous layer was extracted withdichloromethane. The organic layer was dried (Na₂SO₄), filtered, andconcentrated in vacuo. The resulting crude was purified by flashchromatography (SiO₂, 10-50% ethyl acetate in hexanes) to afford thedesired product (0.092 g, 0.18 mmol, 33%).

b) To a stirred solution of the3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexanecarboxylicacid (0.072 g, 0.14 mmol) and HATU (0.11 g, 0.28 mmol) in anhydrousN,N-dimethylforamide (DMF) (1.0 mL) was added a solution of ammonia (0.5M in p-dioxane, 0.5 mL, 0.25 mmol). The reaction mixture was heated at45° C. for 18 h. After cooling to room temperature, the reaction mixturewas quenched with deionized water, extracted with diethyl ether. Theorganic layer was dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was purified by flash chromatography (SiO₂, 10-50%ethyl acetate in hexanes) to afford the title compound as a white solid(0.014 g, 0.028 mmol, 20%). ¹H NMR (400 MHz, CDCl₃) δ 7.37-7.32 (m, 2H),7.16 (dd, J=2.4, 8.4 Hz, 1H), 7.07 (d, J=8.4 Hz, 1H), 6.17 (dd, J=2.4,8.8 Hz, 1H), 5.77 (d, J=4.8 Hz, 1H), 5.43 (br, 1H), 5.29 (br, 1H),4.93-4.88 (m, 1H), 4.65 (d, J=4.8 Hz, 1H), 3.00-2.93 (m, 3H), 2.65-2.63(m, 3H), 2.41-2.34 (m, 1H), 2.23-2.17 (m, 1H), 2.11-2.09 (m, 1H),1.94-1.88 (m, 3H), 1.60 (m, 2H), 1.53 (d, J=6.4 Hz, 3H), 1.46-1.15 (m,4H); MS: (ES) m/z calculated for C₂₅H₃₂Cl₃N₄O [M+H]⁺ 509.2, found 509.3.

Example 16: Synthesis of(3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexyl)methanol(see FIG. 19)

a) To a solution of3-(4-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazin-1-yl)cyclohexanecarboxylicacid (prepared as illustrated in Example 15, 0.077 g, 0.15 mmol) in THF(1 mL) at 0° C. was added sodium borohydride (NaBH₄) (0.017 g, 0.5 mmol)and boron trifluoride diethyl etherate (BF₃.Et₂O) (0.1 mL, 0.8 mmol).The reaction mixture was then stirred at room temperature for 2 h. Thereaction was quenched with aqueous saturated sodium bicarbonate andextracted with dichloromethane. The organic layer was dried (Na₂SO₄),filtered, and concentrated in vacuo. The resulting crude was purified byflash chromatography (SiO₂, 10-40% ethyl acetate in hexanes) to affordthe desired product as a white solid (0.0074 g, 0.015 mmol, 10%). ¹H NMR(400 MHz, CDCl₃) δ 7.37-7.32 (m, 2H), 7.17 (dd, J=2.0, 8.0 Hz, 1H), 7.06(d, J=8.8 Hz, 1H), 6.17 (dd, J=2.8, 8.8 Hz, 1H), 5.78 (d, J=2.4 Hz, 1H),4.90 (dd, J=5.6, 6.4 Hz, 1H), 4.65 (d, J=5.2 Hz, 1H), 3.54 (d, J=7.2 Hz,2H), 3.03-2.99 (m, 3H), 2.59 (m, 3H), 2.39-2.36 (m, 1H), 2.05-1.95 (m,1H), 1.75-1.34 (m, 14H); MS: (ES) m/z calculated for C₂₅H₃₃Cl₃N₃O [M+H]+496.2, found 496.3.

Example 17: Synthesis of2-((R)-1-(2,4-dichlorophenyl)ethylamino)-4-(4-((R)-piperidine-2-carbonyl)piperazin-1-yl)benzonitrile(see FIG. 20)

a) To a solution of 4-bromo-2-fluoro-benzonitrile (2.0 g, 10.0 mmol) and(1R)-1-(2,4-dichlorophenyl)ethanamine (1.9 g, 10.0 mmol) in dimethylsulfoxide (DMSO) (4 mL) was added triethylamine (1.45 g, 14.3 mmol). Thereaction mixture was heated at 120° C. for 20 h. After cooling to roomtemperature, the mixture was diluted with deionized water and a browngummy material was obtained. The aqueous layer was decanted off, and thegum was dissolved in ethyl acetate, washed with deionized water, andbrine. The organic layer was dried (Na₂SO₄), filtered, and concentratedin vacuo. The crude product was used without further purification ((3.4g, 9.2 mmol, 92%).

b) A mixture of(R)-4-bromo-2-(1-(2,4-dichlorophenyl)ethylamino)benzonitrile (0.51 g,1.4 mmol), N-t-butyloxycarbonylpiperazine (0.51 g, 2.8 mmol),tBuDavePhos (0.047 g, 0.13 mmol), and Cs₂CO₃ (1.1 g, 3.5 mmol) in 2:1p-dioxane/triethyl amine(3 mL) was purged with nitrogen for 10 min.Addition of Pd₂(dba)₃ (0.05 g, 0.05 mmol) was followed, and the mixturewas purged with nitrogen for 1 min. The reaction was then heated at 100°C. for 1.5 h. After cooling to room temperature, the mixture wasfiltered and washed with EtOAc (25 mL). The filtrate was concentrated invacuo. The resulting crude mixture was purified by flash chromatography(SiO₂, 0-20% ethyl acetate in hexanes) to afford the coupled product(0.34 g, 0.72 mmol, 52%).

c) Trifluoroacetic acid (0.5 mL) was added to a solution of (R)-t-butyl4-(4-cyano-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazine-1-carboxylate(0.34 g, 0.72 mmol) in dichloromethane (2 mL) at room temperature, andthe solution was stirred for 1 h. Excess solvent was removed in vacuo,and the residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuo.The crude product was used without further purification (0.27 g, 0.72mmol, 100%).

d) To a stirred solution of the(R)-2-(1-(2,4-dichlorophenyl)ethylamino)-4-(piperazin-1-yl)benzonitrile(0.080 g, 0.21 mmol) and (R)-1-(t-butoxycarbonyl)piperdine-2-carboxylicacid (0.049 g, 0.21 mmol) in 1-methyl pyrrolidinone (NMP) (1.0 mL) wasadded HATU (0.085 g, 0.22 mmol) and iPr₂NEt (0.055 g, 0.43 mmol). Thereaction mixture was stirred at room temperature for 2 h, and quenchedwith deionized water. The aqueous layer was extracted with diethyl ether(2×10 mL). The organic layer was dried (Na₂SO₄), filtered, andconcentrated in vacuo. The crude product was purified by flashchromatography (SiO₂, 0-25% ethyl acetate in hexanes) to afford thedesired product (0.095 g, 0.16 mmol, 77%).

e) Trifluoroacetic acid (0.3 mL) was added to a solution of (R)-t-butyl2-(4-(4-cyano-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperazine-1-carbonyl)piperidine-1-carboxylate(0.095 g, 0.16 mmol) in dichloromethane (1 mL) at room temperature, andthe solution was stirred for 1 h. Excess solvent was removed in vacuo,and the residue was diluted with dichloromethane. The organic layer wasneutralized with aqueous saturated sodium bicarbonate solution, and theaqueous layer was further extracted with dichloromethane. The combinedorganic layers were dried (Na₂SO₄), filtered, and concentrated in vacuoto give the title compound as a white solid (0.060 g, 0.12 mmol, 75%).¹H NMR (400 MHz, DMSO-d₆) δ 9.0 (br, 1H), 8.62-8.52 (m, 1H), 7.62-7.50(m, 2H), 7.37 (dd, J=2.2, 8.4 Hz, 1H), 7.05 (d, J=8.8 Hz, 1H), 6.19 (dd,J=2.5, 8.8 Hz, 1H), 5.82 (d, J=2.6 Hz, 1H), 5.68 (br, 1H), 4.86 (br,1H), 3.70-2.80 (m, 9H), 1.98-1.92 (m, 1H), 1.78-1.58 (m, 4H), 1.50-1.40(m, 5H); MS: (ES) m/z calculated for C₂₅H₃₀Cl₂N₅O [M+H]⁺ 486.2, found486.

Example 18: Synthesis of1-(1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)pyrrolidin-3-yl)urea(see FIG. 21)

a) A mixture of (R)-4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyltrifluoromethanesulfonate (Example 2 step d, 5.0 g, 11.1 mmol),4-piperidone ethylene acetal (4.0 g, 27.9 mmol), tBuDavePhos (0.23 g,0.66 mmol), and Cs₂CO₃ (7.2 g, 22.2 mmol) in triethylamine (32 mL) waspurged with nitrogen for 5 min. Addition of Pd₂(dba)₃ (0.31 g, 0.33mmol) was followed, and the mixture was purged with nitrogen for 1 min.The reaction was then heated at 100° C. for 18 h. After cooling to roomtemperature, the mixture was filtered and washed with EtOAc (50 mL). Thefiltrate was concentrated in vacuo, and the resulting crude mixture waspurified by flash chromatography (SiO₂, 0-20% ethyl acetate in hexanes)to afford the coupled product as a viscous oil (1.3 g, 3.0 mmol, 27%).

b) To a solution of2-chloro-N—((R)-1-(2,4-dichlorophenyl)ethyl)-5-(1,4-dioxa-8-azaspiro[4.5]decan-8-yl)aniline(1.3 g, 3.0 mmol) in 5% deionized water in formic acid (5 mL) was heatedat 60° C. for 18 h. The mixture was concentrated in vacuo, and theresidue was partitioned in EtOAc and deionized water. The organic layerwas dried (Na₂SO₄), filtered, and concentrated in vacuo. The crudemixture was purified by flash chromatography (SiO₂, 0-60% ethyl acetatein hexanes) to afford the desired product (0.7 g, 1.76 mmol, 59%).

c) To a stirred solutionof(R)-1-(4-chloro-3-(1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-one(0.13 g, 0.32 mmol) and 3-(t-butoxycarbonylamino)pyrrolidine (0.24 g,1.3 mmol) in dichloroethane (DCE) (2 mL) was added NaBH(OAc)₃ (0.094 g,0.44 mmol). The reaction mixture was heated at 80° C. for 4 h. Thereaction mixture was quenched with aqueous saturated sodium bicarbonateand extracted with dichloromethane. The organic layer was dried(Na₂SO₄), filtered, and concentrated in vacuo. The resulting crude waspurified by flash chromatography (SiO₂, 1-7% methanol indichloromethane) to give t-butyl1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)pyrrolidin-3-ylcarbamate(0.092 g, 0.16 mmol, 31%).

d) Trifluoroacetic acid (0.5 mL) was added to a solution of t-butyl1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)pyrrolidin-3-ylcarbamate(0.092 g, 0.16 mmol) in dichloromethane (1.5 mL) at room temperature,and the solution was stirred for 30 min. The mixture was diluted withdichloromethane, and neutralized with aqueous saturated sodiumbicarbonate solution. The aqueous layer was extracted withdichloromethane, and the organic layer was dried (Na₂SO₄), filtered, andconcentrated in vacuo. The crude product was used without furtherpurification (0.056 g, 0.12 mmol, 75%).

e) To a stirred solution of1-(1-(4-chloro-3-((R)-1-(2,4-dichlorophenyl)ethylamino)phenyl)piperidin-4-yl)pyrrolidin-3-amine(0.056 g, 0.12 mmol) in THF (1 mL) and acetic acid (1 drop) was added asolution of (trimethylsily)isocyanate (0.015 g, 0.13 mmol) in THF (0.5mL). The reaction mixture was stirred at room temperature for 30 min.The reaction mixture was quenched with diluted with aqueous saturatedsodium bicarbonate and extracted with EtOAc. The organic layer was dried(Na₂SO₄), filtered, and concentrated in vacuo. The resulting crude waspurified by flash chromatography (SiO₂, 2-10% methanol indichloromethane) to afford the desired product as a white solid (0.016g, 0.031 mmol, 26%). ¹H NMR (400 MHz, CDCl₃) δ 7.36-7.32 (m, 2H), 7.16(dd, J=2.2, 8.4 Hz, 1H), 7.04 (d, J=8.6 Hz, 1H), 6.82 (br, 1H), 6.16(dd, J=2.5, 8.8 Hz, 1H), 5.77 (br, 1H), 5.23 (br, 1H), 4.87 (q, J=6.6Hz, 1H), 4.64 (d, J=5.2 Hz, 1H), 3.44-3.37 (m, 2H), 3.08-2.82 (m, 3H),2.69-2.40 (m, 4H), 2.25-1.85 (m, 6H), 1.60-1.42 (m, 5H); MS: (ES) m/zcalculated for C₂₄H₃₁Cl₃N₅O [M+H]⁺ 510.2, found 510.

Biological Examples Biological Example 1: Ligand Binding Assay

Ligand binding assay was used to determine the ability of potentialCCR(4) antagonists to block the interaction between CCR(4) and itsligand CCL17 (TARC). CEM cells (ATCC, VA) which naturally express theCCR(4) receptor, were centrifuged and resuspended in assay buffer (20 mMHEPES pH 7.1, 140 mM NaCl, 1 mM CaCl2, 5 mM MgCl2, 0.1% sodium azide andwith 0.1% bovine serum albumin) to a concentration of 5×10{circumflexover ( )}5 cells/mL. Binding assays were set up as follows. First, 0.1mL of cells (5×10⁴ cells/well) was added to the assay plates containingthe compounds, giving a final concentration of ˜2-10 uM each compoundfor screening (or part of a dose response for compound IC50determinations). Then 0.1 mL of 125I labeled TARC(obtained fromPerkinElmer; Waltham, Mass.) diluted in assay buffer to a finalconcentration of ˜50 pM, yielding ˜30,000 cpm per well, was added, theplates sealed and incubated for approximately 3 hours at 25° C. on ashaker platform. Reactions were aspirated onto GF/B glass filterspre-soaked in 0.3% polyethyleneimine (PEI) solution, on a vacuum cellharvester (Packard Instruments; Meriden, Conn.). Scintillation fluid (50uL; Microscint 20, Packard Instruments) was added to each well, theplates were sealed and radioactivity measured in a Top Countscintillation counter (Packard Instruments). Control wells containingeither diluent only (for total counts) or 20 uM compound were used tocalculate the percent of total inhibition for compound. The computerprogram Prism from GraphPad, Inc. (San Diego, Ca) was used to calculateIC50 values. IC50 values are those concentrations required to reduce thebinding of labeled TARC to the receptor by 50%. Compounds in FIG. 22having an IC50 value in the binding assay of less than 100 nM arelabeled (+++); from 100-500 nM are labeled (++); and above 500 nM arelabeled (+).

Biological Example 2

A serum chemotaxis assay was used to determine the efficacy of potentialreceptor antagonists at blocking the migration mediated throughchemokine receptors, such as CCR(4). This assay was routinely performedusing the ChemoTX® microchamber system with a 5-μm pore-sizedpolycarbonate membrane. To begin such an assay, chemokine-receptorexpressing cells (such as CEM cells for the CCR(4) assay) were collectedby centrifugation at 400×g at room temperature, then suspended at 50million/ml in human serum. The compound being tested or an equivalentvolume of its solvent (DMSO) was then added to the cell/serum mixture ata final DMSO concentration of 0. 25% (v/v). Separately, recombinanthuman CCL22 (MDC) was diluted with chemotaxis buffer (HBSS+0.1% BSA),generally spanning a range from 0.01 nM to 500 nM, after which 29 μl ofdiluted chemokine was placed in the lower wells of the ChemoTX® plate.The 5-μm (pore size) polycarbonate membrane was placed onto the plate,and 20 μL of the cell/compound mixture was transferred onto each well ofthe membrane. The plates were incubated at 37° C. for 90 minutes, afterwhich the polycarbonate membranes were removed and 5 μl of theDNA-intercalating agent CyQUANT (Invitrogen, Carlsbad, Calif.) was addedto the lower wells. The amount of fluorescence, corresponding to thenumber of migrated cells, was measured using a Spectrafluor Plus platereader (TECAN, San Jose, Calif.).

Biological Example 3

Compounds of the invention were assessed in the murine model of dermaldelayed type hypersensitivity induced by oxazolone. Briefly, 8-10 weekold BALB/c mice were sensitized topically with a 1% solution ofoxazolone dissolved in ethanol on their shaved abdomens on day 0. On day6 post sensitization mice were dosed orally with either vehicle orincreasing doses of compound 1.127 of the invention immediately prior toand 4 hours following a topical challenge with a 0.5% solution ofoxazolone in ethanol on the right ear. The following day (day 7), earthicknesses were measured using caliper measurements. Animals treatedwith compound had significantly reduced ear swelling compared to vehicletreated controls indicating a compound mediated decrease in oxazoloneinduced dermal hypersensitivity.

Biological Example 4

CCR(4)Compounds of the invention were assessed in the murine model ofallergic asthma. Asthma was induced in 8-10 week old BALB/c mice bysensitizing mice with OVA in Alum adjuvant on days 0 and 10. On day 20mice were challenged with OVA in PBS intranasally to elicit airwayinflammation. Groups of mice were either treated with vehicle, orincreasing doses of compound 1.127 of the invention starting on day 20and lasting until day 23. Animals were subsequently analyzed at day 23after the intranasal OVA challenge for cellular infiltrates inbronchioalveolar lavage (BAL). Mice treated with a compound of theinvention displayed significantly reduced BAL leukocyte numbers relativeto vehicle treated mice at all doses tested.

Biological Example 5

This example describes a procedure to evaluate the efficacy of CCR(4)antagonists for treatment of rheumatoid arthritis. An animal model ofrheumatoid arthritis can be induced in rodents by injecting them withtype II collagen in selected adjuvants. Three series of rodent groupsconsisting of 15 genetically-susceptible mice or rats per group areinjected sub-cutaneously or intra-dermally with type II collagenemulsified in Complete Freund's Adjuvant at days 0 and 21. One series ofrodents additionally receives PBS and Tween 0.5% i.p. at the initialsensitization, and at different dosing schedules thereafter. A secondseries consists of groups of rodents receiving different doses of theCCR(4) antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration at the initial sensitization, and at different dosingschedules thereafter. A third series of rodents, serving as positivecontrol, consists of groups treated with either mouse IL-10 i.p., oranti-TNF antibodies i.p. at the initial sensitization, and at differentdosing schedules thereafter. Animals are monitored from weeks 3 till 8for the development of swollen joints or paws, and graded on a standarddisease severity scale. Disease severity is confirmed by histologicalanalysis of joints.

Biological Example 6

This example describes a procedure to evaluate efficacy of CCR(4)antagonists for treatment of Systemic Lupus Erythematosus (SLE). FemaleNZB/W FI mice spontaneously develop an SLE-like pathology commencing at6 months of age that is characterized by proteinuria, serumautoantibodies, glomerulonephritis, and eventually death. Three seriesof NZB/W FI mouse groups comprising 20 mice per group are tested forefficacy of CCR(4) antagonist as follows: One series of miceadditionally receives phosphate buffered saline (PBS) and Tween 0.5%i.p. soon after weaning, and thereafter at varying dosing schedules. Asecond series consists of groups of mice receiving different doses ofthe CCR(4) antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration soon after weaning, and thereafter at varying dosingschedules. A third series of mice, serving as positive control, consistsof groups treated with anti-IL10 antibodies given soon after weaning,and thereafter at varying dosing schedules. Disease development ismonitored in terms of eventual mortality, kidney histology, serumautoantibody levels, and proteinuria.

Biological Example 7

This example describes a procedure to evaluate efficacy of CCR(4)antagonists for treatment of malignancy. Normal mouse strains can betransplanted with a variety of well-characterized mouse tumor lines,including a mouse thymoma EL4 which has been transfected with OVA toallow easy evaluation of tumor specific antigen responses followingvaccination with OVA. Three series of mouse groups from any of thesetumor models are tested for CCR(4) antagonist efficacy as follows: Oneseries of mice additionally receives PBS and Tween 0.5% i.p. soon aftertumor transplant, and thereafter at varying dosing schedules. A secondseries consists of groups of mice receiving different doses of theCCR(4) antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intramuscularly, orally, or via any other mode ofadministration soon after tumor transplant, and thereafter at varyingdosing schedules. A third series of mice, serving as positive control,consists of groups treated with either anti-IL4 antibodies, anti-IFNgantibodies, IL4, or TNF, given i.p. soon after tumor transplant, andthereafter at varying dosing schedules. Efficacy is monitored via tumorgrowth versus regression. In the case of the OVA-transfected EL4 thymomamodel, cytolytic OVA-specific responses can be measured by stimulatingdraining lymph node cells with OVA in vitro, and measuringantigen-specific cytotoxicity at 72 hours.

Biological Example 8

This example describes procedures to evaluate the efficacy of CCR(4)antagonists in psoriasis. A rodent model of psoriasis can be obtained byintra-venously transferring a population of purified T cells (designatedCD45Rbhi T cells) obtained from the spleens of BALB/c mice intoimmunodeficient recipient CB. 17 scid/scid mice. Mice develop signs ofredness, swelling, and skin lesions resembling those of human psoriasisin their ear, feet and tail by 8 weeks after transfer. Three series ofmouse groups, comprising 10-15 CB.17 scid/scid mice per group, areinjected with purified CD45Rbhi T cells. One series of mice additionallyreceives phosphate buffered saline (PBS) and Tween 0.5% i.p. at theinitial cell transfer, and at different dosing schedules thereafter. Asecond series consists of groups of mice receiving different doses ofthe CCR(4) antagonist given either intra-peritoneally, intra-venously,sub-cutaneously, intra-muscularly, orally, or via any other mode ofadministration at the initial cell transfer, and at different dosingschedules thereafter. A third series of mice, serving as positivecontrol, consists of groups treated with antibodies to either IL-12,IL-4, IFNg, or TNF, or with cytokine IL-10 at the initial cell transfer,and at different dosing schedules thereafter. Animals are monitored fordevelopment of psoriatic-like lesions for 3 months after cell transfer.

Biological Example 9

This example describes a procedure to evaluate the efficacy of CCR(4)antagonists in Inflammatory Bowel Disease (IBD). Several mouse models ofIBD (including Crohn's Disease and Ulcerative Colitis) have beendeveloped. Some of these are spontaneous models occurring in geneticallyengineered transgenic mice that have been depleted of certain cytokinegenes (e.g. IL-IO, or IL-2). Another mouse model of IBD is obtained bytransferring highly purified populations of CD4+T lymphocytes bearing aparticular surface marker phenotype (namely CD45 RB hi) into SCID mice.Three series of mouse groups from anyone ofthese models can be used toevaluate CCR(4) antagonist efficacy as follows. One group of miceadditionally receives PBS and Tween 0.5% i.p. soon after weaning in thecase of the spontaneous models in transgenic mice, or at time of celltransfer into SCID mice and varying dosings thereafter for the celltransfer model. A second series consists of groups of mice receivingdifferent doses of the CCR(4) antagonist given either intraperitoneally,intra-venously, sub-cutaneously, intra-muscularly, orally, or via anyother mode of administration soon after weaning in the case of thespontaneous models in transgenic mice, or at time of cell transfer intoSCID mice and varying dosings thereafter for the cell transfer model. Athird series of mice, serving as positive control, consists of groupstreated with antibodies to either IFNg, or TNF, or with cytokine IL-10soon after weaning in the case of the spontaneous models in transgenicmice, or at time of cell transfer into SCID mice and varying dosingsthereafter for the cell transfer model. Mice are evaluated for 6-8 weeksfor disease development, monitored initially via weight loss and/orprolapsed rectum, and eventually by histological evaluation of theanimals colon and intestinal tract.

Biological Example 10

The mouse RENCA tumor model accurately mimics the progression of humanadult renal cell carcinoma specifically with reference to spontaneousmetastasis to lungs and serves as a model for solid tumors. Balb/c 6-8week old female mice are inoculated with approximately 5e5 RENCA cells(mouse renal adenocarcinoma; ATCC cat # CRL-2947) under the kidneycapsule and kidney tumor growth is observed over 22 days, with lungmetastasis observed as early as day 15. Animals are dosed with eithervehicle or a compound of the invention eg daily subcutaneously, from thetime of tumor implantation to monitor effects on primary growth, or at alater time (eg day 7) to monitor the compound effect on metastasis.Primary tumor areas are measured twice a week using mechanical calipers.Tumor volumes are calculated by the formula v=pab2/6, where a is thelongest diameter and b is the next longest diameter perpendicular to a.A reduction in tumor volume or incidence of metastasis indicatesefficacy of compound in this indication.

It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims.

All publications, patents, and patent applications cited herein arehereby incorporated by reference in their entirety for all purposes.

What is claimed is:
 1. A compound having formula (I):

and any pharmaceutically acceptable salts thereof, wherein R¹ is amember selected from the group consisting of hydrogen, C₁₋₈ alkyl, C₁₋₈haloalkyl, C₁₋₈ hydroxyalkyl, C₃₋₈ cycloalkyl, halogen, —CN, —SO₂Me and—C(O)NH₂; each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen, —CN and C₁₋₈ alkoxy; or two R² groupsattached to adjacent carbon atoms are optionally connected to form a 5-or 6-member ring which is optionally substituted with additional R²groups; R³ is a member selected from the group consisting of hydrogen,methyl and C₁₋₄ haloalkyl; R⁴ is a member selected from the groupconsisting of hydrogen, C₁₋₈ alkyl, C₁₋₈ haloalkyl and C₁₋₈hydroxyalkyl; each of the subscripts n is independently an integer from0 to 3; B is a bond or C(O); Q is a member selected from the groupconsisting of C, CH, N, O, S, S(O) and SO₂; W, X, Y, and Z areindependently selected from the group consisting of C, CH and N, but Qand W are not both N; R⁵ and R⁶ are absent or are members independentlyselected from the group consisting of H, —OH, C₁₋₈ alkyl, C₁₋₈hydroxyalkyl, C₁₋₄ alkoxy-C₁₋₄ alkyl, —C(O)NR^(a)R^(b), C₁₋₈alkylene-C(O)NR^(a)R^(b), —NH—C₁₋₄ alkylene-C(O)NR^(a)R^(b), —C(O)—C₁₋₄alkylene-NR^(a)R^(b), —CO₂H and acid isosteres, C₁₋₈ alkylene-CO₂H andacid isosteres, —N(R^(a))C(O)NR^(a)R^(b), C₁₋₈ alkyleneN(R^(a))C(O)NR^(a)R^(b), —NR^(a)R^(b), C₁₋₈ alkylene-NR^(a)R^(b), C₁₋₈alkoxy, —C(O)OR^(a), C₁₋₈ alkylene-C(O)OR^(a), —CN, —C(O)R^(a),—SO₂R^(a) and —N(R^(a))C(O)R^(b); wherein each R^(a) and R^(b) areindependently selected from the group consisting of hydrogen, C₁₋₈alkyl, C₁₋₈ hydroxyalkyl, C₁₋₈ haloalkyl, and C₁₋₈ alkoxy; and R⁷ isabsent or is selected from the group consisting of H, C₁₋₈ alkyl andC₁₋₈ haloalkyl.
 2. A compound of claim 1, wherein X and Y are not bothN.
 3. A compound of claim 1, wherein R³ is H, and each R² is a memberindependently selected from the group consisting of C₁₋₈ alkyl, C₁₋₈haloalkyl, halogen and —CN.
 4. A compound of claim 3, having formula Ia:


5. A compound of claim 4, wherein X is C or CH.
 6. A compound of claim1, having formula (Ib):

wherein each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen and —CN.
 7. A compound of claim 1, havingformula (Ic):

wherein each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen and —CN; and the subscript n is 0 or 1.8. A compound of claim 7, wherein n is 1, and R⁴ is hydrogen or methyl.9. A compound of claim 1, having formula (Id):

wherein each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen and —CN; and the subscript n is 0 or 1.10. A compound of claim 9, wherein n is 1, and R⁴ is hydrogen or methyl.11. A compound of claim 1, having formula (Ie):

wherein each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen and —CN; and the subscript n is 0 or 1.12. A compound of claim 11, wherein n is 1, and R⁴ is hydrogen ormethyl.
 13. A compound of claim 1, having formula (If):

wherein each R² is a member selected from the group consisting of C₁₋₈alkyl, C₁₋₈ haloalkyl, halogen and —CN.
 14. A compound of any of claims1-6, wherein B is C(O).
 15. A compound of any of claims 1-6, wherein thering having Z as a ring vertex is selected from the group consisting ofpyrrolidine and piperidine.
 16. A compound of any of claims 1-6, whereinthe ring having Z as a ring vertex is selected from the group consistingof pyrrolidin-2-yl and piperidin-2-yl, and at least one of R⁵, R⁶ and R⁷is other than hydrogen.
 17. A compound of any of claims 1-6, wherein Bis a bond.
 18. A compound of any of claims 1-6, wherein B is a bond andthe ring having Z as a ring vertex is selected from the group consistingof pyrrolidine, piperidine and cyclohexane.
 19. A compound of any ofclaims 1-6, wherein B is a bond and the ring having Z as a ring vertexis selected from the group consisting of pyrrolidin-1-yl,pyrrolidin-2-yl, piperidin-1-yl, piperidin-2-yl, piperidin-3-yl andcyclohexane.
 20. A compound of any of claims 1-6, wherein B is a bondand the ring having Z as a ring vertex is selected from the groupconsisting of pyrrolidin-1-yl, pyrrolidin-2-yl, piperidin-1-yl,piperidin-2-yl, piperidin-3-yl and cyclohexane; and at least one of R⁵,R⁶ and R⁷ is other than hydrogen.
 21. A compound of any of claims 1-6,wherein Z is CH or N.
 22. A compound of claim 1, selected from the groupconsisting of:


23. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound of claim
 1. 24. A pharmaceuticalcomposition in accordance with claim 23, comprising a pharmaceuticallyacceptable excipient and a compound of FIG. 22 having ++ or +++activity.
 25. A pharmaceutical composition in accordance with claim 23,comprising a pharmaceutically acceptable excipient and a compound ofFIG. 22 having +++ activity.
 26. A pharmaceutical composition inaccordance with claim 23, comprising a pharmaceutically acceptableexcipient and a compound selected from the group consisting of:


27. A method of treating a disease or condition mediated by CCR(4)signalling, said method comprising administering to a subject in needthereof an efficacious amount of a compound of claim
 1. 28. A method inaccordance with claim 27, wherein said disease or condition is selectedfrom the group consisting of (1) allergic diseases, (2) inflammatorybowel diseases, (3) vaginitis, (4) psoriasis and inflammatorydermatoses, (5) vasculitis, (6) spondyloarthropathies, (7) scleroderma,(8) asthma and respiratory allergic diseases, (9) autoimmune diseases,(10) graft rejection, (11) other diseases in which undesiredinflammatory responses are to be inhibited, and cancer.
 29. A method inaccordance with claim 27, wherein said disease or condition is selectedfrom the group consisting of allergic diseases, psoriasis, atopicdermatitis and asthma.
 30. A method in accordance with claim 27, whereinsaid compound is a compound of FIG. 22 having ++ or +++ activity.
 31. Amethod in accordance with claim 27, wherein said compound is a compoundof FIG. 22 having +++ activity.